CN105993962B - The transplanting acclimation method of strawberry pollen tissue cultural seedlings of free - Google Patents
The transplanting acclimation method of strawberry pollen tissue cultural seedlings of free Download PDFInfo
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- CN105993962B CN105993962B CN201610603534.2A CN201610603534A CN105993962B CN 105993962 B CN105993962 B CN 105993962B CN 201610603534 A CN201610603534 A CN 201610603534A CN 105993962 B CN105993962 B CN 105993962B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- 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
- A01G31/00—Soilless cultivation, e.g. hydroponics
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Abstract
The invention discloses the transplanting acclimation methods of strawberry anther tissue cultural seedlings of free, include the following steps：1）Pollen collection and cleaning and sterilizing, 2）Induce differentiation into seedling, 3）Seedling is fast numerous with taking root, and 4）It emerges and washes seedling, 5）Transplanting, 6）Domestication.Beneficial effects of the present invention are：Method provided by the invention, seedling need not be practiced, directly clean culture medium can be transplanted, plastic canopy is built on seedbed, closing effectively controls temperature in arch membrane, soil humidity, light application time and intensity after three days, up to 100%, the probability for obtaining disease-free strain greatly improves the survival rate of pollen tissue cultural seedlings of free；Simultaneously, strawberry pollen virus elimination rate can be up to 100%, virus checker can not be had to, shorten the period from detoxification to large-scale production seedling, effectively reduce production cost and labour cost, productivity effect is increased substantially, provides large batch of high-quality virus-elimination seedlings, technical support is provided for the sustainable industrialization sound development of strawberry.
Description
Technical field
The present invention relates to plant tissue culture seedling-raising technique fields, and in particular to the transplanting domestication side of strawberry pollen tissue cultural seedlings of free
Method.
Background technology
The test tube seedling of pollen plant mainly carries out heterotrophism metabolism on culture medium by the sugar in culture medium, and is trying
Humidity and temperature are all more stable in pipe, once moving into soil, great variety occurs for physiological ecological environment, often due to test tube seedling
This variation cannot be adapted to quickly and causes large quantities of death, particularly transplanted and changed frequently just when natural temperature height, move on schedule
Plant is difficult to survive.Generally use at present is that there are two stage hardening processes for bottle seedling：First, tissue-culture container seedling moves on to outdoor in natural light
Lower placement 2 d~3 d practice seedling；Second is that open the d of 3 d of lid hardening~5 again, the result is that short, the children that changes to autotrophy ambient time by heterotrophism
Seedling is there are no suitable, then opens lid, and pollution rate, which rises rapidly, can reach 30%~60%, substantially reduces the resistance of seedling, transplants
Survival rate significantly declines afterwards.
Invention content
The purpose of the present invention aiming at it is above-mentioned in the prior art the defects of, provide the shifting of strawberry pollen tissue cultural seedlings of free
Plant acclimation method.
To achieve these goals, technical solution provided by the invention is：The transplanting domestication of strawberry pollen tissue cultural seedlings of free
Method includes the following steps：
1）The materials and cleaning and sterilizing of strawberry pollen：
Using Field strawberries pollen development to the anther of the mid-late uninucleate stage of 4mm~6mm sizes, pollen mid-late uninucleate stage
Strawberry bud form is：Calyx does not open, and bud is slightly longer than corolla or corolla just shows money or valuables one carries unintentionally, corolla white or pale green and not loose
It moves, anther is micro- yellow and enriches；The fresh anther taken is placed on after being stored for 24 hours in 2 DEG C~4 DEG C, with tap water by anther surface
It rinses well；Flushed anther is moved on superclean bench and is put into sterile beaker, first with 75% 30 s of alcohol disinfecting~40s,
Aseptic water washing 2 times~3 times, then the min of 4 min~5 is sterilized with 0.1% mercuric chloride solution, with the glass bar to sterilize along one
Direction is gently agitated for, and then with aseptic water washing 4 times~5 times, 1 min, is blotted rapidly with filter paper every time, obtains sterile anther；
2）The induction of anther callus and cauline leaf differentiation：
Aseptically, the bud sterilized is fixed on the big rubber for having been subjected to sterilizing with sterile needle；With having gone out
The scalpel and dissecting needle of bacterium peel off bud, take out anther, the anther of taking-up is inoculated into equipped with callus inducing medium
On, callus inducing medium composition is：1.5 mg/L of MS+ BA~0.10 mg/L of 2.5mg/L+NAA~0.25mg/L
+ 3% sucrose+6g/L~8g/L agar, pH value 5.8；As 2 mm of diameter~3 mm of callus, it is transferred to differential medium
In, differential medium composition is：+ 3% sucrose in 0.08 mg/L of MS+ BA~0.8 mg/L of 0.12mg/L+IBA~1.2mg/L
+ 6g/L~8g/L agar, pH value 5.8；Bud is cut into small pieces when forming apparent Multiple Buds, is inoculated in Multiplying culture of growing thickly
It is proliferated on base, proliferated culture medium of growing thickly composition is：1.2 mg/L of MS+BA~1.8mg/L+NAA, 0.08 mg/L~
0.12mg/+3% sucrose+6g/L~8g/L agar L, pH value 5.8；
3）The fast numerous and culture of rootage of tissue cultural seedlings of free：
By step 2）Obtained detoxic seedling is put into 25d~35d in proliferated culture medium and grows up to bottle seedling of growing thickly；Proliferated culture medium is
Add in the MS culture mediums of 6- benzyl aminoadenines 0.19mg/L~0.21mg/L；High 3cm, robust growth are inoculated in without offspring
On MS culture mediums, after 2 weeks, seedling base portion grows radial white rootlet, and every plant takes root after 5~7,3 weeks, 1 cm of root growth
~1.5cm, for transplanting；
4）Matrix prepares before transplanting：
The composition of matrix is：According to weight ratio turf：Perlite：Vermiculite=3：1：1；
The matrix for mixing, having soaked is packed into the hole tray in 32,50 caves or the battalion of diameter 9cm~12cm by the 1d before transplanting seedlings
It supports in alms bowl, it is light to press；It is irrigated with 50% 1000 times of liquid of wettable powder of carbendazim, when transplanting seedlings within second day, water content of substrate 70%
~80 % are suitble to transplant seedlings；
5）Bottle outlet washes seedling classification：
It plays before seedling all to be sterilized operation instrument used with 75 % alcohol swabs, the tissue culture bottle taken root directly is removed from culturing room
Seedling from bottle is sorted out tissue-cultured seedling to washing chamber, with diameter 1cm tools, is placed in 25 DEG C~28 DEG C of clear water and cleans the training of root
Base is supported, root system, tender shoots and blade is not hurt, then by size be classified seedling, and separate no offspring；
6）Transplanting：
The tissue-cultured seedling of clean classification is transported to seedling nursery solarium and is so transplanted by the same day, with ready tool in hole tray
Matrix on insert hole, seedling root is gently put into hole, is gently covered, is slightly compacted；It is gently sprayed water, rinsed with sprayer after having moved
Matrix in seedling leaves so that root system contacted with matrix it is closer, in favor of root growth；For no offspring, it is suitable to be first stained with
Concentration：The root-inducing powder or root-growing agent of NAA 0.05mg/L~0.1mg/L or IBA 0. 8mg/L~1.0mg/L, then transplant or move
After plant pouring root is carried out with the root-inducing powder of a concentration of 800 times of liquid；
7）Domestication：
Plastics Small plastic shed must be built on seedbed by having transplanted the same day, sealed every morning after 3d or lifted film 30min at dusk, from
The 10% of membrane area is taken off in side, starts to tame tissue-cultured seedling；Then film-uncovering time increases 10min daily, and taking off membrane area increases by 10%；Depending on
Temperature height and compound seedling medium water content in environment suitably increase the illumination ventilated time, can be thrown off after the d of 10d~15 thin
Film；It needs to carry out shading management after transplanting, the preceding 2d sunshade net shadings 60%~70% after transplanting, 10d~14d later is gradual
Light is seen, until not shading；The tissue culture shoot survival percent of domestication is up to can move to the nutritive cube of a diameter of 10cm after 100%, 30d or educate
It is cultivated in seedbed.
Further, the transplanting acclimation method of above-mentioned strawberry pollen tissue cultural seedlings of free, the step 3）Tissue cultural seedlings of free
Before fast numerous and culture of rootage, enzyme linked immunosorbent assay may be used, i.e. ELISA method carries out viral diagnosis.
Further, the transplanting acclimation method of above-mentioned strawberry pollen tissue cultural seedlings of free, the step 2）In, antherderived callus
The induction of tissue and cauline leaf differential growth condition are：23 DEG C~25 DEG C, 2500 lx of intensity of illumination~3000lx of temperature, illumination 14
H/d~16h/d, 20 d of incubation time~30d.
Further, the transplanting acclimation method of above-mentioned strawberry pollen tissue cultural seedlings of free, the step 3）In, detoxication and tissue culture
Miao Kuaifan and the growth conditions of culture of rootage are：23 DEG C~25 DEG C, 2000 lx of intensity of illumination~3000lx, 10 h/ of illumination of temperature
D~12h/d, fast numerous and two stage 50 d of incubation time~60d of taking root.
Further, the transplanting acclimation method of above-mentioned strawberry pollen tissue cultural seedlings of free, the step 6）In, transplanting condition
For：15 DEG C~25 DEG C of temperature.
Further, the transplanting acclimation method of above-mentioned strawberry pollen tissue cultural seedlings of free, the step 7）In, acclimation conditions
For：20 DEG C~38 DEG C of temperature, 20 d of incubation time~30d.
Beneficial effects of the present invention are：The transplanting acclimation method of strawberry pollen tissue cultural seedlings of free provided by the invention uses
Before transplanting need not to the white silk seedling process of bottle seedling, directly from the Strawberry Plantlets bottle seedling that tissue culture room removal is taken root to washing chamber,
Directly open bottle cap seedling taking, with the culture medium of 25 DEG C~28 DEG C of warm water cleaning root, the same day transport to seedling nursery solarium so into
Row transplanting, the transplanting same day build plastic canopy on seedbed, and closing effectively controls temperature in arch membrane, soil humidity, light after three days
According to time and intensity, the survival rate of pollen tissue cultural seedlings of free is up to 100%, using pollen cultures technology compared with Shoot Tip Culture and Re Chu
The probability that reason detoxification obtains disease-free strain greatly improves；Meanwhile pollen detoxification do not have to virus checker, shorten from detoxification to
In the period of large-scale production seedling, production cost and labour cost are effectively reduced, increase substantially productivity effect, provided large quantities of
The high-quality virus-elimination seedlings of amount provide technical support for the sustainable industrialization sound development of strawberry.
Specific embodiment
Preparation source used in the present invention：
Carbendazim：Producer：Anhui Huaxing Chemical Co., Ltd.；Agriculture chemical registration card number：PD85150-19；Production permit
Card number：XK13-003-00126;Product standard card number：HG3290-2000；Lot number：On 01 20th, 2014；
Root-growing agent（Powder）：
The producer of NAA：Shanghai Blue Season Technology Development Co., Ltd Chembase, lot number：Lot No.B0013K0321017；
The producer of IBA：Shanghai Blue Season Technology Development Co., Ltd's Chembase registration numbers：CAS 133-32-4 lot numbers：
141212。
Embodiment 1：
The transplanting acclimation method of strawberry pollen tissue cultural seedlings of free, includes the following steps：
1）The materials and cleaning and sterilizing of strawberry pollen：
Using Field strawberries pollen development to the anther of the mid-late uninucleate stage of 4mm~6mm sizes, pollen mid-late uninucleate stage
Strawberry bud form is：Calyx does not open, and bud is slightly longer than corolla or corolla just shows money or valuables one carries unintentionally, corolla white or pale green and not loose
It moves, anther is micro- yellow and enriches；The fresh anther taken is placed on after being stored for 24 hours in 2 DEG C~4 DEG C, with tap water by anther surface
It rinses well；Flushed anther is moved on superclean bench and is put into sterile beaker, first with 75% 30 s of alcohol disinfecting~40s,
Aseptic water washing 2 times~3 times, then the min of 4 min~5 is sterilized with 0.1% mercuric chloride solution, with the glass bar to sterilize along one
Direction is gently agitated for, and then with aseptic water washing 4 times~5 times, 1 min, is blotted rapidly with filter paper every time, obtains sterile anther；
2）The induction of anther callus and cauline leaf differentiation：
Aseptically, the bud sterilized is fixed on the big rubber for having been subjected to sterilizing with sterile needle；With having gone out
The scalpel and dissecting needle of bacterium peel off bud, take out anther, the anther of taking-up is inoculated into equipped with callus inducing medium
On, callus inducing medium composition is：1.5 mg/L of MS+ BA~0.10 mg/L of 2.5mg/L+NAA~0.25mg/L+
3% sucrose+6g/L~8g/L agar, pH value 5.8；As diameter 2mm~3mm of callus, it is transferred to differential medium MS+
In 0.08 mg/L of BA~0.8 mg/L of 0.12mg/L+IBA~1.2mg/L+3% sucrose+6g/L~8g/L agar, pH value is
5.8；Bud is cut into small pieces when forming apparent Multiple Buds, is inoculated on proliferated culture medium of growing thickly and is proliferated, proliferation of growing thickly
Culture medium composition is 1.2 mg/L of MS+BA~0.08 mg/L of 1.8mg/L+NAA~0.12mg/L+3% sucrose+6g/L~8g/L
Agar, pH value 5.8；
The induction of anther callus and cauline leaf differential growth condition are：23 DEG C~25 DEG C of temperature, 2500 lx of intensity of illumination
~3000lx, 14 h/d of illumination~16h/d, 20 d of incubation time~30d；
3）The fast numerous and culture of rootage of tissue cultural seedlings of free：
Before this step, enzyme linked immunosorbent assay can be used, i.e. ELISA method carries out viral diagnosis, can not also detect；
By step 2）Obtained detoxic seedling is put into 25 d in proliferated culture medium~35d and grows up to bottle seedling of growing thickly；Proliferated culture medium
To add in the MS culture mediums of 6- benzyl aminoadenines 0.19mg/L~0.21mg/L；By high 3cm, robust growth without offspring be inoculated with
In on MS culture mediums, after 2 weeks, seedling base portion grows radial white rootlet, and every plant takes root after 5~7,3 weeks, root growth 1
Cm~1.5cm, for transplanting；
Numerous and culture of rootage growth conditions is tissue cultural seedlings of free soon：23 DEG C~25 DEG C of temperature, 2000 lx of intensity of illumination~
3000lx, 10 h/d of illumination~12h/d, fast numerous and two stage 50 d of incubation time~60d of taking root；
4）Matrix prepares before transplanting：
Matrix formulations：Turf；Perlite：Vermiculite=3:1:1；
1 day before transplanting seedlings, the matrix for mixing, having soaked is packed into the hole tray in 32,50 caves or diameter 9cm~12cm
It is light to press in nutritive cube；It is irrigated with 50% 1000 times of liquid of wettable powder of carbendazim, when transplanting seedlings within second day, water content of substrate is
70%~80 % is suitble to transplant seedlings；
5）Bottle outlet washes seedling classification：
It plays before seedling all to be sterilized operation instrument used with 75 % alcohol swabs, the tissue culture bottle taken root directly is removed from culturing room
Seedling from bottle is sorted out tissue-cultured seedling to washing chamber, with diameter 1cm tools, is placed in 25 DEG C~28 DEG C of clear water and cleans the training of root
Base is supported, root system, tender shoots and blade is not hurt, then by size be classified seedling, and separate no offspring；
6）Transplanting：
The tissue-cultured seedling of clean classification is transported to seedling nursery solarium and is so transplanted by the same day, with ready tool in hole tray
Matrix on insert hole, seedling root is gently put into hole, is gently covered, is slightly compacted；It is gently sprayed water, rinsed with sprayer after having moved
Matrix in seedling leaves so that root system contacted with matrix it is closer, in favor of root growth；For no offspring, it is suitable to be first stained with
Concentration：The root-inducing powder or root-growing agent of NAA 0.05mg/L~0.1mg/L or IBA 0. 8mg/L~1.0mg/L, then transplant or move
After plant pouring root is carried out with the root-inducing powder of a concentration of 800 times of liquid；Transplanting condition is：15 DEG C~25 DEG C of temperature；
7）Domestication：
Plastics Small plastic shed must be built on seedbed by having transplanted the same day, sealed every morning after 3d or lifted film 30min at dusk, from
The 10% of membrane area is taken off in side, starts to tame tissue-cultured seedling；Then film-uncovering time increases 10min daily, and taking off membrane area increases by 10%；Depending on
Temperature height and compound seedling medium water content in environment suitably increase the illumination ventilated time, can be thrown off after the d of 10d~15 thin
Film；It needs to carry out shading management after transplanting, the preceding 2d sunshade net shadings 60%~70% after transplanting, 10d~14d later is gradual
Light is seen, until not shading；The tissue culture shoot survival percent of domestication is up to can move to the nutritive cube of a diameter of 10cm after 100%, 30d or educate
It is cultivated in seedbed；Acclimation conditions are：20 DEG C~38 DEG C of temperature, 20 d of incubation time~30d.
Finally it should be noted that：The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
To modify to the technical solution recorded in foregoing embodiments or carry out equivalent replacement to which part technical characteristic.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in the present invention's
Within protection domain.
Claims (5)
1. the transplanting acclimation method of strawberry pollen tissue cultural seedlings of free, which is characterized in that include the following steps：
1）The materials and cleaning and sterilizing of strawberry pollen：
Using Field strawberries pollen development to the anther of the mid-late uninucleate stage of 4mm~6mm sizes, the strawberry of pollen mid-late uninucleate stage
Bud form is：Calyx does not open, and bud is slightly longer than corolla or corolla just shows money or valuables one carries unintentionally, corolla white or pale green and does not loosen, spends
Medicine is micro- yellow and enriches；The fresh anther taken is placed on after being stored for 24 hours in 2 DEG C~4 DEG C, with tap water by anther surface washing
Totally；Flushed anther is moved on superclean bench and is put into sterile beaker, first with the s of 75% alcohol disinfecting, 30 s~40, nothing
Bacterium water rinses 2 times~3 times, then sterilizes the min of 4 min~5 with 0.1% mercuric chloride solution, with the glass bar to sterilize along a side
To being gently agitated for, then with aseptic water washing 4 times~5 times, 1 minute every time, blotted with filter paper rapidly, obtain sterile anther；
2）The induction of anther callus and cauline leaf differentiation：
Aseptically, the bud sterilized is fixed on the big rubber of sterilized mistake with sterile needle；With sterilized
Scalpel and dissecting needle peel off bud, take out anther, the anther of taking-up are inoculated into equipped on callus inducing medium, more
Injured tissue inducing culture forms：1.5 mg/L of MS+ BA~2.5,0.10 mg/L of mg/L+NAA~0.25mg/L+3% sugarcanes
The g/L agar of+6 g/L of sugar~8, pH value 5.8；As 2 mm of diameter~3 mm of callus, it is transferred in differential medium,
Differential medium forms：The mg/L+3% sucrose+6 of 0.08 mg/L of MS+ BA~0.8 mg/L of 0.12mg/L+IBA~1.2
Bud is cut into small pieces when forming apparent Multiple Buds, is inoculated in Multiplying culture of growing thickly by the g/L agar of g/L~8, pH value 5.8
It is proliferated on base, proliferated culture medium of growing thickly composition is：1.2 mg/L of MS+BA~1.8 mg/L+NAA, 0.08 mg/L~
The g/L agar of+6 g/L of 0.12 mg/L+3% sucrose~8, pH value 5.8；
3）The fast numerous and culture of rootage of tissue cultural seedlings of free：
Before the fast numerous and culture of rootage of tissue cultural seedlings of free, using enzyme linked immunosorbent assay, i.e. ELISA method carries out viral diagnosis；
Again by step 2）Obtained detoxic seedling is put into 25d~35d in proliferated culture medium and grows up to bottle seedling of growing thickly；Proliferated culture medium is adds in 6-
The MS culture mediums of the mg/L of 0.19 mg/L of benzyl aminoadenine~0.21；By high 3cm, robust growth without offspring be inoculated in MS training
It supports on base, after 2 weeks, seedling base portion grows radial white rootlet, and every plant takes root after 5~7,3 weeks, 1 cm~1. of root growth
5cm, for transplanting；
4）Matrix prepares before transplanting：
The composition of matrix is：According to weight ratio turf：Perlite：Vermiculite=3：1：1；
1 day before transplanting seedlings, the matrix for mixing, having soaked is packed into the nutritive cube of the 9~12cm of hole tray or diameter in 32,50 caves
In, it is light to press；It is irrigated with 50% 1000 times of liquid of wettable powder of carbendazim, when transplanting seedlings within second day, water content of substrate is 70%~80
% is suitble to transplant seedlings；
5）Bottle outlet washes seedling classification：
Before seedling is played all to be sterilized operation instrument used with 75 % alcohol swabs, directly from culturing room's tissue-culture container seedling for taking root of removal to
Washing chamber from bottle sorts out tissue-cultured seedling with diameter 1cm tools, is placed in 25 DEG C~28 DEG C of clear water and cleans the culture of root
Base not hurt root system, tender shoots and blade, then by size be classified seedling, and separate no offspring；
6）Transplanting：
The tissue-cultured seedling of clean classification is transported to seedling nursery solarium and is then transplanted by the same day, with ready tool in hole tray
Matrix on insert hole, seedling root is gently put into hole, is gently covered, is slightly compacted；It is gently sprayed water, rinsed with sprayer after having moved
Matrix in seedling leaves so that root system contacted with matrix it is closer, in favor of root growth；For no offspring, it is suitable to be first stained with
Concentration：The root-inducing powder or root-growing agent of the mg/L of 0.8 mg/L of NAA 0.05mg/L~0.1mg/L or IBA~1.0, then transplant or
After transplanting pouring root is carried out with the root-inducing powder of a concentration of 800 times of liquid；
7）Domestication：
Plastics Small plastic shed must be built on seedbed by having transplanted the same day, sealed every morning after 3d or lifted film 30min at dusk, from side
The 10% of membrane area is taken off, starts to tame tissue-cultured seedling；Then film-uncovering time increases 10min daily, and taking off membrane area increases by 10%；Depending on environment
In temperature height and compound seedling medium water content suitably increase the illumination ventilated time, can throw off film after the d of 10d~15；It moves
It needing to carry out shading management, the preceding 2d sunshade net shadings 60%~70% after transplanting after plant, 10 d~14d later is gradually shown in light,
Until not shading；The tissue culture shoot survival percent of domestication up to the nutritive cube or seedling bed that a diameter of 10cm can be moved to after 100%, 30d into
Row is cultivated.
2. the transplanting acclimation method of strawberry pollen tissue cultural seedlings of free according to claim 1, which is characterized in that the step
2）In, the induction of anther callus and cauline leaf differential growth condition are：23 DEG C~25 DEG C of temperature, 2500 lx of intensity of illumination~
3000lx, 14 h/d of illumination~16h/d, 20 d of incubation time~30d.
3. the transplanting acclimation method of strawberry pollen tissue cultural seedlings of free according to claim 1, which is characterized in that the step
3）In, fast numerous and culture of rootage the growth conditions of tissue cultural seedlings of free is：23 DEG C~25 DEG C of temperature, 2000 lx of intensity of illumination~
3000lx, 10 h/d of illumination~12h/d, fast numerous and two stage 50 d of incubation time~60d of taking root.
4. the transplanting acclimation method of strawberry pollen tissue cultural seedlings of free according to claim 1, which is characterized in that the step
6）In, transplanting condition is：15 DEG C~25 DEG C of temperature.
5. the transplanting acclimation method of strawberry pollen tissue cultural seedlings of free according to claim 1, which is characterized in that the step
7）In, acclimation conditions are：20 DEG C~38 DEG C of temperature, 20 d of incubation time~30d.
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| CN101049090A (en) * | 2007-05-22 | 2007-10-10 | 南京农业大学 | Method for carrying out taking off poison and quick breeding by using strawberry anther |
| CN103858758A (en) * | 2012-12-12 | 2014-06-18 | 东港市草莓研究所 | Strawberry seedling tissue culture rapid propagation technology |
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