CN105993962A - Transplanting and acclimatizating method of virus-free tissue culture seedlings of strawberry pollen - Google Patents
Transplanting and acclimatizating method of virus-free tissue culture seedlings of strawberry pollen Download PDFInfo
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- CN105993962A CN105993962A CN201610603534.2A CN201610603534A CN105993962A CN 105993962 A CN105993962 A CN 105993962A CN 201610603534 A CN201610603534 A CN 201610603534A CN 105993962 A CN105993962 A CN 105993962A
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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
Abstract
The invention discloses a transplanting and acclimatizating method of virus-free tissue culture seedlings of strawberry pollen. The method comprises the following steps: (1) collecting, washing and sterilizing pollen; (2) inducing differentiation of pollen into seedlings; (3) rapidly propagating seedlings and rooting; (4) emerging seedlings and washing the seedlings; (5) transplanting; and (6) acclimatizating. The method is capable of transplanting by directly washing a culture medium without the need of hardening the seedlings; a plastic arched shed is built on a seedling bed; subsequently, the plastic arched shed is sealed for three days; then the temperature, the substrate humidity, the illumination time and the illumination intensity in an arched film are effectively controlled. Through the method, the survival rate of the virus-free tissue culture seedlings of the pollen can reach 100%; the probability of obtaining virus-free seedlings is greatly improved; meanwhile, the virus removal rate of the strawberry pollen can reach 100%; the method is free of virus detection process; the period from virus removal to large-scale production of the seedlings is shortened; the production cost and the labor cost are effectively reduced; the production benefit is greatly improved; large batches of high-quality virus-free seedlings are provided; the technical support is supplied to the sustainable and healthy industrialization development of strawberry.
Description
Technical field
The present invention relates to plant tissue culture seedling-raising technique field, be specifically related to the transplanting domestication side of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free
Method.
Background technology
The test tube Seedling of pollen plant mainly carries out heterotrophism metabolism by the sugar in culture medium in culture medium, and in examination
In pipe, humidity and temperature are the most more stable, once move into soil, physiological ecological environment generation great variety, often due to test tube Seedling
Can not quickly adapt to this change and cause large quantities of death, particularly transplant just when natural temperature height change frequently, move on schedule
Cultivation is difficult to survive.The most commonly used is that a bottle Seedling has two stage seedling exercising processes: one is that tissue-culture container seedling moves on to outdoor at natural light
Lower placement 2 d～3 d practices Seedling；Two is to open lid seedling exercising 3 d～5 d again, and result is that to be changed to autotrophy ambient time by heterotrophism short, children
Seedling is not also suitable, then opens lid, and pollution rate rises rapidly and can reach 30%～60%, is substantially reduced the resistance of seedling, transplants
Rear survival rate significantly declines.
Summary of the invention
The purpose of the present invention is aiming at above-mentioned defect of the prior art, it is provided that the shifting of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free
Plant acclimation method.
To achieve these goals, the technical scheme that the present invention provides is: the transplanting domestication of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free
Method, comprises the following steps:
1) the drawing materials and cleaning and sterilizing of Fructus Fragariae Ananssae pollen:
Use Field strawberries pollen development to the flower pesticide of the mid-late uninucleate stage of 4mm～6mm size, the Fructus Fragariae Ananssae of pollen mid-late uninucleate stage
Alabastrum form is: calyx does not opens, alabastrum slightly longer than corolla or corolla just shows money or valuables one carries unintentionally, corolla white or pale green and loosen, flower
The micro-Huang of medicine and enrich；After the fresh flower pesticide taked is placed in 2 DEG C～4 DEG C storage 24h, with tap water, flower pesticide surface washing is done
Only；Flushed flower pesticide moves into be put in sterile beaker, first with 75% alcohol disinfecting 30 s～40s, sterilized water on superclean bench
Rinse 2 times～3 times, then sterilize 4 min～5 min with the mercuric chloride solution of 0.1%, light along a direction with the Glass rod of sterilizing
Light agitation, then uses aseptic water washing 4 times～5 times, and each 1 min blots with filter paper rapidly, obtains aseptic flower pesticide；
2) induction and the stem and leaf of anther callus breaks up:
Aseptically, by sterile needle, the alabastrum sterilized is fixed on the big rubber of sterilizing；With sterilized
Dissecting knife and dissecting needle strip off alabastrum, take out flower pesticide, is inoculated into by the flower pesticide of taking-up equipped with on callus inducing medium, heals
Injured tissue inducing culture consists of: MS+ BA 1.5 mg/L～2.5mg/L+NAA 0.10 mg/L～0.25mg/L+3% sugarcane
Sugar+6g/L～8g/L agar, pH value is 5.8；As diameter 2 mm～3 mm of callus, proceed in division culture medium, point
Change culture medium to consist of :+3% sucrose+6g/L in MS+ BA 0.08 mg/L～0.12mg/L+IBA 0.8 mg/L～1.2mg/L
～8g/L agar, pH value is 5.8；When forming obvious Multiple Buds, bud is cut into small pieces, is inoculated in and grows thickly on proliferated culture medium
Breeding, proliferated culture medium of growing thickly consists of: MS+BA 1.2 mg/L～1.8mg/L+NAA 0.08 mg/L～0.12mg/+
3% sucrose+6g/L～8g/L agar L, pH value is 5.8；
3) tissue cultural seedlings of free is the most numerous and root culture:
By step 2) detoxic seedling that obtains puts into 25d～35d in proliferated culture medium and grows up to a bottle Seedling of growing thickly；Proliferated culture medium is for adding
The MS culture medium of 6-benzyl aminoadenine 0.19mg/L～0.21mg/L；By high 3cm, robust growth without root be inoculated in MS training
Supporting on base, after 2 weeks, Seedling base portion grows radial white rootlet, and every strain is taken root 5～7, after 3 weeks, root growth 1 cm～1.
5cm, is used for transplanting；
4) before transplanting, substrate prepares:
Consisting of of substrate: according to the weight ratio peat composed of rotten mosses: perlite: Vermiculitum=3:1:1；
1d before transplanting seedlings, by mixing, the substrate of moistening load hole tray or the nutritive cube of diameter 9cm～12cm in 32,50 caves
In, gently press；1000 times of liquid of wettability powder of carbendazim with 50% irrigate, and when within second day, transplanting seedlings, water content of substrate is 70%～80
%, is i.e. suitable for transplanting seedlings；
5) bottle outlet washes Seedling classification:
With 75 % alcohol swabs, operation instrument used is all sterilized before lifting, directly from culturing room's tissue-culture container seedling of taking root of removal to
Laundry room, sorts out tissue cultured seedling with diameter 1cm instrument from bottle, is placed in the clear water of 25 DEG C～28 DEG C the cultivation cleaning root
Base, does not hurt root system, tender shoots and blade, the most by size by seedling classification, and separates without root；
6) transplant:
The same day tissue cultured seedling of clean classification was transported to seedling nursery solarium so transplant, with ready instrument at the base of hole tray
Insert hole in matter, seedling root is put in hole gently, covers gently, be slightly compacted；Gently spray water with aerosol apparatus after having moved, rinse seedling
Substrate on blade so that root system contacts tightr with substrate, is beneficial to root growth；For without root, first it is stained with suitable concentration:
NAA 0.05mg/L～0.1mg/L or the root-inducing powder of IBA 0. 8mg/L～1.0mg/L or root-growing agent, then transplant, or after transplanting
Pouring root is carried out with the root-inducing powder that concentration is 800 times of liquid；
7) domestication:
Having transplanted and must build plastics Small plastic shed the same day on seedbed, after sealing 3d, every morning or dusk lift film 30min, from side
Take off the 10% of membrane area, start to tame tissue cultured seedling；Film-uncovering time every day increases 10min subsequently, and taking off membrane area increases by 10%；Depending on environment
In temperature height and seedling medium water content suitably increase illumination ventilated time, 10d～15 d after can throw off thin film；Move
Needing after planting to carry out shading management, the front 2d sunshade net shading 60%～70% after transplanting, 10d～14d afterwards is gradually shown in light,
Until not shading；The tissue cultured seedling survival rate 100% of domestication, the nutritive cube or the seedling bed that just can be moved to a diameter of 10cm after 30d enter
Row is cultivated.
Further, the transplanting acclimation method of above-mentioned Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, described step 3) tissue cultural seedlings of free
Before the most numerous and root culture, enzyme linked immunosorbent assay, i.e. ELISA method can be used to carry out Viral diagnosis.
Further, the transplanting acclimation method of above-mentioned Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, described step 2) in, antherderived callus
Tissue induction and stem and leaf differential growth condition be: temperature 23 DEG C～25 DEG C, intensity of illumination 2500 lx～3000lx, illumination 14
H/d～16h/d, incubation time 20 d～30d.
Further, the transplanting acclimation method of above-mentioned Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, in described step 3), detoxication and tissue culture
The growth conditions of Miao Kuaifan and root culture is: temperature 23 DEG C～25 DEG C, intensity of illumination 2000 lx～3000lx, illumination 10 h/
D～12h/d, the most numerous and two stage incubation time 50 d～60d of taking root.
Further, the transplanting acclimation method of above-mentioned Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, in described step 6), transplanting condition
For: temperature 15 DEG C～25 DEG C.
Further, the transplanting acclimation method of above-mentioned Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, in described step 7), acclimation conditions
For: temperature 20 DEG C～38 DEG C, incubation time 20 d～30d.
The invention have the benefit that the transplanting acclimation method of the Fructus Fragariae Ananssae pollen tissue cultural seedlings of free that the present invention provides, use
Before transplanting, bottle Seedling need not be practiced Seedling process, the Strawberry Plantlets bottle Seedling directly taken root from tissue culture room removal to laundry room,
Directly open bottle cap seedling taking, by the culture medium of the warm water cleaning root of 25 DEG C～28 DEG C, transported to seedling nursery solarium the same day and so enter
Row is transplanted, and transplants and built plastic canopy on seedbed the same day, effectively controls temperature, soil humidity, light in arch film after closing three days
According to time and intensity, the survival rate of pollen tissue cultural seedlings of free, up to 100%, uses pollen cultures technology relatively Shoot Tip Culture and Re Chu
Reason detoxification obtains the probability of anosis strain and is greatly improved；Meanwhile, pollen detoxification without virus checker, shorten from detoxification to
In the cycle of large-scale production seedling, effectively reduce production cost and labour cost, increase substantially productivity effect, it is provided that be large quantities of
The high-quality virus-elimination seedlings of amount, develops in a healthy way for the sustainable industrialization of Fructus Fragariae Ananssae and provides technical support.
Detailed description of the invention
Preparation source used by the present invention:
Carbendazim: producer: Anhui Huaxing Chemical Co., Ltd.；Agriculture chemical registration card number: PD85150-19；Production licence
Number: XK13-003-00126;Product standard card number: HG3290-2000；Lot number: on 01 20th, 2014；
Root-growing agent (powder):
The producer of NAA: upper sea blue season development in science and technology company limited Chembase, lot number: Lot No.B0013K0321017；
The producer of IBA: upper sea blue season development in science and technology company limited Chembase registration number: CAS 133-32-4 lot number:
141212。
Embodiment 1:
The transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, comprises the following steps:
1) the drawing materials and cleaning and sterilizing of Fructus Fragariae Ananssae pollen:
Use Field strawberries pollen development to the flower pesticide of the mid-late uninucleate stage of 4mm～6mm size, the Fructus Fragariae Ananssae of pollen mid-late uninucleate stage
Alabastrum form is: calyx does not opens, alabastrum slightly longer than corolla or corolla just shows money or valuables one carries unintentionally, corolla white or pale green and loosen, flower
The micro-Huang of medicine and enrich；After the fresh flower pesticide taked is placed in 2 DEG C～4 DEG C storage 24h, with tap water, flower pesticide surface washing is done
Only；Flushed flower pesticide moves into be put in sterile beaker, first with 75% alcohol disinfecting 30 s～40s, sterilized water on superclean bench
Rinse 2 times～3 times, then sterilize 4 min～5 min with the mercuric chloride solution of 0.1%, light along a direction with the Glass rod of sterilizing
Light agitation, then uses aseptic water washing 4 times～5 times, and each 1 min blots with filter paper rapidly, obtains aseptic flower pesticide；
2) induction and the stem and leaf of anther callus breaks up:
Aseptically, by sterile needle, the alabastrum sterilized is fixed on the big rubber of sterilizing；With sterilized
Dissecting knife and dissecting needle strip off alabastrum, take out flower pesticide, is inoculated into by the flower pesticide of taking-up equipped with on callus inducing medium, heals
Injured tissue inducing culture consists of: MS+ BA 1.5 mg/L～2.5mg/L+NAA 0.10 mg/L～0.25mg/L+3% sugarcane
Sugar+6g/L～8g/L agar, pH value is 5.8；As the diameter 2mm～3mm of callus, proceed to division culture medium MS+ BA
In 0.08 mg/L～0.12mg/L+IBA 0.8 mg/L～1.2mg/L+3% sucrose+6g/L～8g/L agar, pH value is 5.8；
When forming obvious Multiple Buds, bud is cut into small pieces, is inoculated on proliferated culture medium of growing thickly and breeds, enrichment culture of growing thickly
Basis set become MS+BA 1.2 mg/L～1.8mg/L+NAA 0.08 mg/L～0.12mg/L+3% sucrose+6g/L～8g/L fine jade
Fat, pH value is 5.8；
Induction and the stem and leaf differential growth condition of anther callus be: temperature 23 DEG C～25 DEG C, intensity of illumination 2500 lx～
3000lx, illumination 14 h/d～16h/d, incubation time 20 d～30d；
3) tissue cultural seedlings of free is the most numerous and root culture:
Before this step, enzyme linked immunosorbent assay, i.e. ELISA method can be used to carry out Viral diagnosis, it is possible to do not detect；
By step 2) detoxic seedling that obtains puts into 25 d～35d in proliferated culture medium and grows up to a bottle Seedling of growing thickly；Proliferated culture medium is for adding
Enter the MS culture medium of 6-benzyl aminoadenine 0.19mg/L～0.21mg/L；By high 3cm, robust growth be inoculated in MS without root
In culture medium, after 2 weeks, Seedling base portion grow radial white rootlet, every strain takes root 5～7, after 3 weeks, root growth 1 cm～
1.5cm, is used for transplanting；
The most numerous and root culture the growth conditions of tissue cultural seedlings of free is: temperature 23 DEG C～25 DEG C, intensity of illumination 2000 lx～
3000lx, illumination 10 h/d～12h/d, the most numerous and two stage incubation time 50 d～60d of taking root；
4) before transplanting, substrate prepares:
Matrix formulations: the peat composed of rotten mosses；Perlite: Vermiculitum=3:1:1；
Before transplanting seedlings 1 day, by mixing, the substrate of moistening load hole tray or the nutrition of diameter 9cm～12cm in 32,50 caves
In alms bowl, gently press；1000 times of liquid of wettability powder of carbendazim with 50% irrigate, when within second day, transplanting seedlings, water content of substrate be 70%～
80 %, are i.e. suitable for transplanting seedlings；
5) bottle outlet washes Seedling classification:
With 75 % alcohol swabs, operation instrument used is all sterilized before lifting, directly from culturing room's tissue-culture container seedling of taking root of removal to
Laundry room, sorts out tissue cultured seedling with diameter 1cm instrument from bottle, is placed in the clear water of 25 DEG C～28 DEG C the cultivation cleaning root
Base, does not hurt root system, tender shoots and blade, the most by size by seedling classification, and separates without root；
6) transplant:
The same day tissue cultured seedling of clean classification was transported to seedling nursery solarium so transplant, with ready instrument at the base of hole tray
Insert hole in matter, seedling root is put in hole gently, covers gently, be slightly compacted；Gently spray water with aerosol apparatus after having moved, rinse seedling
Substrate on blade so that root system contacts tightr with substrate, is beneficial to root growth；For without root, first it is stained with suitable concentration:
NAA 0.05mg/L～0.1mg/L or the root-inducing powder of IBA 0. 8mg/L～1.0mg/L or root-growing agent, then transplant, or after transplanting
Pouring root is carried out with the root-inducing powder that concentration is 800 times of liquid；Transplanting condition is: temperature 15 DEG C～25 DEG C；
7) domestication:
Having transplanted and must build plastics Small plastic shed the same day on seedbed, after sealing 3d, every morning or dusk lift film 30min, from side
Take off the 10% of membrane area, start to tame tissue cultured seedling；Film-uncovering time every day increases 10min subsequently, and taking off membrane area increases by 10%；Depending on environment
In temperature height and seedling medium water content suitably increase illumination ventilated time, 10d～15 d after can throw off thin film；Move
Needing after planting to carry out shading management, the front 2d sunshade net shading 60%～70% after transplanting, 10d～14d afterwards is gradually shown in light,
Until not shading；The tissue cultured seedling survival rate 100% of domestication, the nutritive cube or the seedling bed that just can be moved to a diameter of 10cm after 30d enter
Row is cultivated；Acclimation conditions is: temperature 20 DEG C～38 DEG C, incubation time 20 d～30d.
Finally it is noted that the foregoing is only the preferred embodiments of the present invention, it is not limited to the present invention,
Although being described in detail the present invention with reference to previous embodiment, for a person skilled in the art, it still may be used
So that the technical scheme described in foregoing embodiments to be modified, or wherein portion of techniques feature is carried out equivalent.
All within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, should be included in the present invention's
Within protection domain.
Claims (6)
1. the transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free, it is characterised in that comprise the following steps:
1) the drawing materials and cleaning and sterilizing of Fructus Fragariae Ananssae pollen:
Use Field strawberries pollen development to the flower pesticide of the mid-late uninucleate stage of 4mm～6mm size, the Fructus Fragariae Ananssae of pollen mid-late uninucleate stage
Alabastrum form is: calyx does not opens, alabastrum slightly longer than corolla or corolla just shows money or valuables one carries unintentionally, corolla white or pale green and loosen, flower
The micro-Huang of medicine and enrich；After the fresh flower pesticide taked is placed in 2 DEG C～4 DEG C storage 24h, with tap water by flower pesticide surface washing
Totally；Flushed flower pesticide moves into be put in sterile beaker, first by 75% alcohol disinfecting 30 s～40 s, nothing on superclean bench
Bacterium water rinses 2 times～3 times, then sterilizes 4 min～5 min with the mercuric chloride solution of 0.1%, with the Glass rod of sterilizing along a side
To being gently agitated for, then use aseptic water washing 4 times～5 times, each 1 minute, blot with filter paper rapidly, obtain aseptic flower pesticide；
2) induction and the stem and leaf of anther callus breaks up:
Aseptically, by sterile needle, the alabastrum sterilized is fixed on the big rubber of sterilizing；With sterilized
Dissecting knife and dissecting needle strip off alabastrum, take out flower pesticide, is inoculated into by the flower pesticide of taking-up equipped with on callus inducing medium, heals
Injured tissue inducing culture consists of: MS+ BA 1.5 mg/L～2.5 mg/L+NAA 0.10 mg/L～0.25mg/L+3% sugarcanes
Sugar+6 g/L～8 g/L agar, pH value is 5.8；As diameter 2 mm～3 mm of callus, proceed in division culture medium,
Division culture medium consists of: MS+ BA 0.08 mg/L～0.12mg/L+IBA 0.8 mg/L～1.2 mg/L+3% sucrose+6
G/L～8 g/L agar, pH value is 5.8, is cut into small pieces by bud when forming obvious Multiple Buds, is inoculated in enrichment culture of growing thickly
Breeding on base, proliferated culture medium of growing thickly consists of: MS+BA 1.2 mg/L～1.8 mg/L+NAA 0.08 mg/L～
0.12 mg/L+3% sucrose+6 g/L～8 g/L agar, pH value is 5.8；
3) tissue cultural seedlings of free is the most numerous and root culture:
By step 2) detoxic seedling that obtains puts into 25d～35d in proliferated culture medium and grows up to a bottle Seedling of growing thickly；Proliferated culture medium is for adding
The MS culture medium of 6-benzyl aminoadenine 0.19 mg/L～0.21 mg/L；By high 3cm, robust growth be inoculated in MS without root
In culture medium, after 2 weeks, Seedling base portion grow radial white rootlet, every strain takes root 5～7, after 3 weeks, root growth 1 cm～
1.5cm, is used for transplanting；
4) before transplanting, substrate prepares:
Consisting of of substrate: according to the weight ratio peat composed of rotten mosses: perlite: Vermiculitum=3:1:1；
Before transplanting seedlings 1 day, by mixing, the substrate of moistening load hole tray or the nutritive cube of diameter 9～12cm in 32,50 caves
In, gently press；1000 times of liquid of wettability powder of carbendazim with 50% irrigate, and when within second day, transplanting seedlings, water content of substrate is 70%～80
%, is i.e. suitable for transplanting seedlings；
5) bottle outlet washes Seedling classification:
With 75 % alcohol swabs, operation instrument used is all sterilized before lifting, directly from culturing room's tissue-culture container seedling of taking root of removal to
Laundry room, sorts out tissue cultured seedling with diameter 1cm instrument from bottle, is placed in the clear water of 25 DEG C～28 DEG C the cultivation cleaning root
Base, does not hurt root system, tender shoots and blade, the most by size by seedling classification, and separates without root；
6) transplant:
The same day tissue cultured seedling of clean classification was transported to seedling nursery solarium so transplant, with ready instrument at the base of hole tray
Insert hole in matter, seedling root is put in hole gently, covers gently, be slightly compacted；Gently spray water with aerosol apparatus after having moved, rinse seedling
Substrate on blade so that root system contacts tightr with substrate, is beneficial to root growth；For without root, first it is stained with suitable concentration:
NAA 0.05mg/L～0.1mg/L or IBA 0.8 mg/L～the root-inducing powder of 1.0 mg/L or root-growing agent, then transplant, or after transplanting
Pouring root is carried out with the root-inducing powder that concentration is 800 times of liquid；
7) domestication:
Having transplanted and must build plastics Small plastic shed the same day on seedbed, after sealing 3d, every morning or dusk lift film 30min, from side
Take off the 10% of membrane area, start to tame tissue cultured seedling；Film-uncovering time every day increases 10min subsequently, and taking off membrane area increases by 10%；Depending on environment
In temperature height and seedling medium water content suitably increase illumination ventilated time, 10d～15 d after can throw off thin film；Move
Needing after planting to carry out shading management, the front 2d sunshade net shading 60%～70% after transplanting, 10 d～14d afterwards are gradually shown in light,
Until not shading；The tissue cultured seedling survival rate 100% of domestication, the nutritive cube or the seedling bed that just can be moved to a diameter of 10cm after 30d enter
Row is cultivated.
The transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free the most according to claim 1, it is characterised in that described step
3) tissue cultural seedlings of free is the most numerous and before root culture, uses enzyme linked immunosorbent assay, i.e. ELISA method to carry out Viral diagnosis.
The transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free the most according to claim 1, it is characterised in that described step
2) in, induction and the stem and leaf differential growth condition of anther callus be: temperature 23 DEG C～25 DEG C, intensity of illumination 2500 lx～
3000lx, illumination 14 h/d～16h/d, incubation time 20 d～30d.
The transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free the most according to claim 1, it is characterised in that described step
3), in, the most numerous and root culture the growth conditions of tissue cultural seedlings of free is: temperature 23 DEG C～25 DEG C, intensity of illumination 2000 lx～
3000lx, illumination 10 h/d～12h/d, the most numerous and two stage incubation time 50 d～60d of taking root.
The transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free the most according to claim 1, it is characterised in that described step
6) in, transplanting condition is: temperature 15 DEG C～25 DEG C.
The transplanting acclimation method of Fructus Fragariae Ananssae pollen tissue cultural seedlings of free the most according to claim 1, it is characterised in that described step
7) in, acclimation conditions is: temperature 20 DEG C～38 DEG C, incubation time 20 d～30d.
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| CN109964755A (en) * | 2019-05-15 | 2019-07-05 | 十堰市经济作物研究所 | A method of improving Original Strawberry kind shoot survival percent and quality |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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2016
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (5)
| Title |
|---|
| 任建宏等: "草莓‘拉松7号’花药培养研究", 《安徽农业科学》 * |
| 胡婷婷等: ""贝吉佳"草莓脱毒苗生根及驯化研究", 《安徽农业科学》 * |
| 郭艳等: "红颜草莓脱毒苗炼苗移栽技术研究", 《现代园艺》 * |
| 陈文胜: "草莓引种试验与繁殖推广", 《中国优秀学位论文全文数据库 农业科技辑》 * |
| 高公泓等: "草莓离体无性繁殖的研究", 《西北植物学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109964755A (en) * | 2019-05-15 | 2019-07-05 | 十堰市经济作物研究所 | A method of improving Original Strawberry kind shoot survival percent and quality |
| CN109964755B (en) * | 2019-05-15 | 2021-03-23 | 十堰市经济作物研究所 | Method for improving survival rate and quality of strawberry stock seedlings |
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| CN105993962B (en) | 2018-07-03 |
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