CN110150152A - A kind of method of tissue cultivating strawberry - Google Patents

A kind of method of tissue cultivating strawberry Download PDF

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Publication number
CN110150152A
CN110150152A CN201910570392.8A CN201910570392A CN110150152A CN 110150152 A CN110150152 A CN 110150152A CN 201910570392 A CN201910570392 A CN 201910570392A CN 110150152 A CN110150152 A CN 110150152A
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culture
culture medium
strawberry
medium
root
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秦燕
朱艺萌
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Chengdu Academy of Agriculture and Forestry Sciences
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Chengdu Academy of Agriculture and Forestry Sciences
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001Culture apparatus for tissue culture
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008Methods for regeneration to complete plants

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  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention discloses a kind of methods of tissue cultivating strawberry, including materials sterilization and prepare culture medium, detoxification and transplanting and other steps, the detoxification, strawberry stolon stem apex is first taken to induce through initial culture base into adventitious bud, the adventitious bud is induced through subculture medium into plantlet group again, then the plantlet is rolled into a ball again and is induced through root media into test tube virus-elimination seedlings, preliminary culture medium, subculture medium is the culture medium of inducing strawberry cell differentiation, root media is the Nutrient medium that inducing strawberry is taken root, the Nutrient medium is made of basal nutrient base MS and nutritional additive.The tissue-cultured seedling that the method comes out strawberry stolon Shoot Tip Culture, well developed root system is not easy to pollute, and can directly transplant, and does not need hardening;This method detoxification is easy to operate, and reproduction speed is fast, at low cost, and yield is high, is suitble to scale breeding.

Description

A kind of method of tissue cultivating strawberry
Technical field
The present invention relates to strawberry seedling breeding field, in particular to a kind of method of tissue cultivating strawberry.
Background technique
Strawberry is rosaceae Fragaria perennial root herbaceos perennial, is worldwide berry, cultivated area and yield Grape is only second in the production of world's berry fruits.The adaptable wide, cultivation of strawberry is easy, result is early, yield is high, income The advantages that good is a kind of higher natural fruit of economic value, therefore all parts of the country are all introduced a fine variety in positive, but due to long-term Stolon vegetative propagation, the reason of susceptible viral invades and variety deterioration is made to thereby result in low output, low output mainly I The long-term continuous cropping of state strawberry, the more seldom update using detoxic seedling of strawberry germ.The virosis of invasion strawberry at present, mainly There are four types of, i.e., Strawberry mottle virus, strawberry crinkle virus, strawberry be light and yellow edge is malicious, strawberry veinbanding virus, and certain kind of berries virosis is serious The yield for influencing strawberry, generally results in underproduction 30%-50%.It is to prevent and treat virosis and improve having for survival rate using tissue cultures Effect approach is also to speed up the technical way of the new stock breeding promotion rate of strawberry and Virus-free seedcane production.
Summary of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of method of tissue cultivating strawberry, this method The tissue-cultured seedling well developed root system turned out, it is not easy to pollute, it can directly transplant, not need hardening;Detoxification is convenient, and reproduction speed is fast, at This is low, and this method induction success rate is high and can safely and fast breed excellent strawberry detoxification tissue culture seedling.
The purpose of the present invention is achieved through the following technical solutions: a kind of method of tissue cultivating strawberry, including with Lower step:
S1: culture medium is sterilized and prepared to materials;
Materials sterilisation step: choosing strawberry stolon 3~5cm of stem apex, first rinses 0.5~1.5h with flowing water and cleans silt, Stem apex is immersed in 10~15min of disinfection in mercuric chloride solution again, it is then clean with aseptic water washing again, ethanol disinfection 30~ 45s finally uses aseptic water washing again, repeats the step 2~3 times;
Prepare culture medium: preparation initial culture base, subculture medium, root media add in three kinds of culture mediums Enter 7-11g/L agar, forms film solid media;
The initial culture base includes: MS culture medium, 0.1~1.0mg/L 6BA;
The subculture medium includes: MS culture medium, 0.1~1.0mg/L 6BA;
The root media includes: MS culture medium, 0.05~0.2mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strips out stem apex growth Point, places it in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, transfer Into the subculture medium of another Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3 It is cut into individual plantlet in superclean bench when~5cm, is transferred in the root media of root induction, is trained Form test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, will It is transplanted into the hole tray for filling sterilization matrix, In Shade to be trained tissue-cultured seedling.
The formula of the MS culture medium are as follows:
A great number of elements: KNO3 1.9g/L、NH4NO3 1.65g/L、MgSO4·7H2O 0.37g/L、KH2PO40.17g/L、 CaCl2·2H2O 0.44g/。
Microelement: MnSO4·4H2O 22.3mg/L、ZnSO4·7H2O 8.6mg/L、H3BO3 6.2mg/L、KI 0.83mg/L、Na2MoO4·7H2O 0.25mg/L、CuSO4. 5H2O4 0.025mg/L、CoCL2. 6H2O0.025mg/。
Molysite: Na2-EDTA 37.3mg/L、FeSO4. 4H2O 27.8mg/L。
Organic matter: glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, Tyiamine Hd element 0.1mg/L, niacin 0.5mg/L, Creatine 100mg/L.
The formula of three kinds of culture mediums is respectively as follows:
Initial culture base includes: MS culture medium, 0.5mg/L 6BA;
Subculture medium includes: MS culture medium, 0.5mg/L 6BA;
Root media includes: MS culture medium, 0.1mg/L IBA.
In the step S1, the ethyl alcohol is 75% medical ethanol.
In the step S1, the concentration of the mercuric chloride solution is 0.1%.
In the step S1, the sterile water is ultrapure water.
It is 25 DEG C in temperature in the step S2, Initial culture is carried out under dark conditions, incubation time is 30~35d; It is 25 DEG C in temperature, culture of rootage is carried out under day light irradiation time 12h, incubation time is 30~35d.
In the step S3, the sterilization matrix is coco bran: perlite=2:1 mixture.
The beneficial effects of the present invention are:
1) method of the invention can turn out the tissue-cultured seedling of well developed root system, and the tissue-cultured seedling is not easy to pollute, can directly transplant, Hardening is not needed, excellent strawberry detoxification tissue culture seedling can be safely and fast bred;And its whole process detoxification is convenient, reproduction speed Fastly, at low cost, yield is high.
2) formula for improving culture medium has been increased separately proportion in basal nutrient base MS and has been closed using solid medium Suitable nutritional additive 6BA and IBA, being respectively intended to can be with the differentiation of inducing strawberry cell and inducing strawberry tissue cultures seedling root Development elongation, induction it is high-efficient.
3) this method is using stem apex of crawling as explant, has materials convenient, and virus is few, is inoculated with high-efficient, and heredity is steady The advantages that determining.
Specific embodiment
Below in conjunction with embodiment, technical solution of the present invention is clearly and completely described, it is clear that described Embodiment is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field Technical staff's every other embodiment obtained under the premise of not making the creative labor belongs to what the present invention protected Range.
Embodiment 1
A kind of method of tissue cultivating strawberry, comprising the following steps:
S1: culture medium is sterilized and prepared to materials;
Materials sterilisation step: choosing strawberry stolon stem apex 5cm, first rinses 1h with flowing water and cleans silt, then stem apex is soaked Bubble sterilizes 10min in mercuric chloride solution, and then clean with aseptic water washing again, ethanol disinfection 30s is finally rushed with sterile water again It washes, repeats the step 2~3 times;
Prepare culture medium: preparation initial culture base, subculture medium, root media add in three kinds of culture mediums Enter 9g/L agar, forms film solid media;
The initial culture base includes: MS culture medium, 0.1mg/L 6BA;
The subculture medium includes: MS culture medium, 0.1mg/L 6BA;
The root media includes: MS culture medium, 0.05mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strips out stem apex growth Point, places it in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, transfer Into the subculture medium of another Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3 It is cut into individual plantlet in superclean bench when~5cm, is transferred in the root media of root induction, is trained Form test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, will It is transplanted into the hole tray for filling sterilization matrix, In Shade to be trained tissue-cultured seedling.
In the step S1, the ethyl alcohol is 75% medical ethanol, and the concentration of the mercuric chloride solution is 0.1%, institute The sterile water stated is ultrapure water.
In the step S2, it is 25 DEG C in temperature, carries out Initial culture, incubation time 30d under dark conditions;In temperature Degree is 25 DEG C, carries out culture of rootage, incubation time 30d under day light irradiation time 12h.
In the step S3, the sterilization matrix is coco bran: perlite=2:1 mixture.
Embodiment 2
A kind of method of tissue cultivating strawberry, comprising the following steps:
S1: culture medium is sterilized and prepared to materials;
It draws materials sterilisation step: choosing strawberry stolon stem apex 5cm, first rinse 1.5h with flowing water and clean silt, then by stem apex It is immersed in mercuric chloride solution and sterilizes 15min, then clean with aseptic water washing again, ethanol disinfection 45s finally uses sterile water again It rinses, repeats the step 2~3 times;
Prepare culture medium: preparation initial culture base, subculture medium, root media add in three kinds of culture mediums Enter 11g/L agar, forms film solid media;
The initial culture base includes: MS culture medium, 0.5mg/L 6BA;
The subculture medium includes: MS culture medium, 0.5mg/L 6BA;
The root media includes: MS culture medium, 0.2mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strips out stem apex growth Point, places it in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, transfer Into the subculture medium of another Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3 It is cut into individual plantlet in superclean bench when~5cm, is transferred in the root media of root induction, is trained Form test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, will It is transplanted into the hole tray for filling sterilization matrix, In Shade to be trained tissue-cultured seedling.
In the step S1, the ethyl alcohol is 75% medical ethanol, and the concentration of the mercuric chloride solution is 0.1%, institute The sterile water stated is ultrapure water.
In the step S2, it is 25 DEG C in temperature, carries out Initial culture, incubation time 35d under dark conditions;In temperature Degree is 25 DEG C, carries out culture of rootage, incubation time 35d under day light irradiation time 12h.
In the step S3, the sterilization matrix is coco bran: perlite=2:1 mixture.
Embodiment 3
A kind of method of tissue cultivating strawberry, comprising the following steps:
S1: culture medium is sterilized and prepared to materials;
It draws materials sterilisation step: choosing strawberry stolon stem apex 4cm, first rinse 0.5h with flowing water and clean silt, then by stem apex It is immersed in mercuric chloride solution and sterilizes 13min, then clean with aseptic water washing again, ethanol disinfection 35s finally uses sterile water again It rinses, repeats the step 2~3 times;
Prepare culture medium: preparation initial culture base, subculture medium, root media add in three kinds of culture mediums Enter 7g/L agar, forms film solid media;
The initial culture base includes: MS culture medium, 0.8mg/L 6BA;
The subculture medium includes: MS culture medium, 0.4mg/L 6BA;
The root media includes: MS culture medium, 0.15mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strips out stem apex growth Point, places it in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, transfer Into the subculture medium of another Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3 It is cut into individual plantlet in superclean bench when~5cm, is transferred in the root media of root induction, is trained Form test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, will It is transplanted into the hole tray for filling sterilization matrix, In Shade to be trained tissue-cultured seedling.
In the step S1, the ethyl alcohol is 75% medical ethanol, and the concentration of the mercuric chloride solution is 0.1%, institute The sterile water stated is ultrapure water.
In the step S2, it is 25 DEG C in temperature, carries out Initial culture, incubation time 32d under dark conditions;In temperature Degree is 25 DEG C, carries out culture of rootage, incubation time 34d under day light irradiation time 12h.
In the step S3, the sterilization matrix is coco bran: perlite=2:1 mixture.
Embodiment 4
A kind of method of tissue cultivating strawberry, comprising the following steps:
S1: culture medium is sterilized and prepared to materials;
It draws materials sterilisation step: choosing strawberry stolon stem apex 3cm, first rinse 1.2h with flowing water and clean silt, then by stem apex It is immersed in mercuric chloride solution and sterilizes 10min, then clean with aseptic water washing again, ethanol disinfection 45s finally uses sterile water again It rinses, repeats the step 2~3 times;
Prepare culture medium: preparation initial culture base, subculture medium, root media add in three kinds of culture mediums Enter 8g/L agar, forms film solid media;
The initial culture base includes: MS culture medium, 0.4mg/L 6BA;
The subculture medium includes: MS culture medium, 0.9mg/L 6BA;
The root media includes: MS culture medium, 0.12mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strips out stem apex growth Point, places it in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, transfer Into the subculture medium of another Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3 It is cut into individual plantlet in superclean bench when~5cm, is transferred in the root media of root induction, is trained Form test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, will It is transplanted into the hole tray for filling sterilization matrix, In Shade to be trained tissue-cultured seedling.
In the step S1, the ethyl alcohol is 75% medical ethanol, and the concentration of the mercuric chloride solution is 0.1%, institute The sterile water stated is ultrapure water.
In the step S2, it is 25 DEG C in temperature, carries out Initial culture, incubation time 35d under dark conditions;In temperature Degree is 25 DEG C, carries out culture of rootage, incubation time 30d under day light irradiation time 12h.
In the step S3, the sterilization matrix is coco bran: perlite=2:1 mixture.
Embodiment 5
A kind of method of tissue cultivating strawberry, comprising the following steps:
S1: culture medium is sterilized and prepared to materials;
It draws materials sterilisation step: choosing strawberry stolon stem apex 3cm, first rinse 1.3h with flowing water and clean silt, then by stem apex It is immersed in mercuric chloride solution and sterilizes 15min, then clean with aseptic water washing again, ethanol disinfection 40s finally uses sterile water again It rinses, repeats the step 2~3 times;
Prepare culture medium: preparation initial culture base, subculture medium, root media add in three kinds of culture mediums Enter 11g/L agar, forms film solid media;
The initial culture base includes: MS culture medium, 0.75mg/L 6BA;
The subculture medium includes: MS culture medium, 0.35mg/L 6BA;
The root media includes: MS culture medium, 0.18mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strips out stem apex growth Point, places it in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, transfer Into the subculture medium of another Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3 It is cut into individual plantlet in superclean bench when~5cm, is transferred in the root media of root induction, is trained Form test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, will It is transplanted into the hole tray for filling sterilization matrix, In Shade to be trained tissue-cultured seedling.
In the step S1, the ethyl alcohol is 75% medical ethanol, and the concentration of the mercuric chloride solution is 0.1%, institute The sterile water stated is ultrapure water.
In the step S2, it is 25 DEG C in temperature, carries out Initial culture, incubation time 32d under dark conditions;In temperature Degree is 25 DEG C, carries out culture of rootage, incubation time 35d under day light irradiation time 12h.
In the step S3, the sterilization matrix is coco bran: perlite=2:1 mixture.
The formula of MS culture medium in above 5 embodiments are as follows:
A great number of elements: KNO3 1.9g/L、NH4NO3 1.65g/L、MgSO4·7H2O 0.37g/L、KH2PO40.17g/L、 CaCl2·2H2O 0.44g/。
Microelement: MnSO4·4H2O 22.3mg/L、ZnSO4·7H2O 8.6mg/L、H3BO3 6.2mg/L、KI 0.83mg/L、Na2MoO4·7H2O 0.25mg/L、CuSO4. 5H2O4 0.025mg/L、CoCL2. 6H2O0.025mg/。
Molysite: Na2-EDTA 37.3mg/L、FeSO4. 4H2O 27.8mg/L。
Organic matter: glycine 2.0mg/L, puridoxine hydrochloride 0.5mg/L, Tyiamine Hd element 0.1mg/L, niacin 0.5mg/L, Creatine 100mg/L.
The method of tissue cultivating strawberry of the invention makes to cultivate high-quality tissue culture after Dongying City strawberry research institute uses Seedling achieves breakthrough progress, realizes large-scale industrialized production, and the annual output of current high-quality tissue cultural seedlings of free reaches 32000000 plants or more, Strawberry tissue culture quick-breeding method of the invention walks the high-quality tissue-cultured seedling of strawberry research institute, Donggang City really Laboratory out realizes large-scale industrialized production.Year can be ten thousand plants of tissue-cultured seedling 2000-3000 of the high-quality detoxification of production and sales at present. By the tissue-cultured seedling of the strawberry of this technical solution production, all from materials, contact size, subculture number and induction success rate etc. Very big improvement, and the step of eliminating previous hardening have been arrived, the rate of induction is not only increased and has also assured kind Purity.The strawberry studies Plantlets of Strawberry produced, and field, which produces, similar " terrible fruit " or " flower never occurs in cultivation And it is not firm " etc. variation phenomenons.The tissue-cultured seedling obtained with stem tip tissue culture detoxification compared with conventional seedling significantly grow it is excellent Gesture and effect of increasing production.The strawberry detoxic seedling of different cultivars, than normal young plant, plant plant height increases by 33%~35%, and leaf area increases 60% or so, stolon quantity increases 90% or so, improves the yield and quality of plant, wherein increase yield up to 46%~ 50%.
The strawberry research institute is carried out continuously strawberry detoxic seedling and conventional seedling (object of reference) 3 years (2015,2016,2017) Hothouse production comparative test and the large-scale numerous seedling of strawberry seed farm are tested, the results show that detoxic seedling increases by 35% than conventional seedling plant height; Leaf area increases by 30%;Single fruit weight increases by 36%;Fruit yield increases by 37%;Fruit soluble solids content increases by 12%; Breeding coefficient improves 45%.Change it can be seen that turning out the Strawberry Plantlets come through method of the invention and taking off seedling than conventional seedling economical character Good, economic characters are excellent, and reproductive capacity is strong, increase production and improve productivity obvious.
Multiple groups have been done to the root growth situation and survival rate of the tissue-cultured seedling cultivated by this method by the strawberry research institute It repeats to test, test result is as shown in table 1.0:
Table 1.0
Above-mentioned first group to the 5th group of experimental subjects is the tissue-cultured seedling cultivated by method of the invention, and the 6th group For conventional young plant, the tissue-cultured seedling well developed root system that method of the invention is turned out it can be seen from table 1.0, high survival rate.This method The Plantlets of Strawberry of cultivation, it is not easy to pollute, it can directly transplant, not need hardening;Detoxification is easy to operate, and reproduction speed is fast, cost Low, yield is high, is suitble to scale breeding.
The above is only a preferred embodiment of the present invention, it should be understood that the present invention is not limited to described herein Form should not be regarded as an exclusion of other examples, and can be used for other combinations, modifications, and environments, and can be at this In the text contemplated scope, modifications can be made through the above teachings or related fields of technology or knowledge.And those skilled in the art institute into Capable modifications and changes do not depart from the spirit and scope of the present invention, then all should be in the protection scope of appended claims of the present invention It is interior.

Claims (7)

1. a kind of method of tissue cultivating strawberry, it is characterised in that: the following steps are included:
S1: culture medium is sterilized and prepared to materials;
It draws materials sterilisation step: choosing strawberry stolon 3~5cm of stem apex, first rinse 0.5 ~ 1.5h with flowing water and clean silt, then by stem Point is immersed in disinfection 10 ~ 15min, then clean with aseptic water washing again, 30 ~ 45s of ethanol disinfection in mercuric chloride solution, finally again With aseptic water washing, repeat the step 2~3 times;
Prepare culture medium: 7- is added in three kinds of culture mediums in preparation initial culture base, subculture medium, root media 11g/L agar forms film solid media;
The initial culture base includes: MS culture medium, 0.1~1.0mg/L 6BA;
The subculture medium includes: MS culture medium, 0.1~1.0mg/L 6BA;
The root media includes: MS culture medium, 0.05~0.2mg/L IBA;
S2: detoxification: first by the aseptic paper suck dry moisture of the stem apex after sterilization, dissecting under the microscope, strip out shoot tip meristem, will It is placed in the initial culture base of Cell differentiation inducing activity, is sealed with sealed membrane, when adventitious bud it is long to 2~4cm when, be transferred to another In the subculture medium of Cell differentiation inducing activity, induce the adventitious buds differentiation at plantlet group, to plantlet regimental commander to 3~5cm When it is cut into individual plantlet in superclean bench, transfer in the root media of root induction, be trained Test tube seedling, i.e. virus-elimination seedlings;
S3: transplanting: test tube seedling is taken out from root media, and the culture medium of test tube seedling root is cleaned with detoxification water, is moved It plants into the hole tray for filling sterilization matrix, it is In Shade to be trained tissue-cultured seedling.
2. the method for tissue cultivating strawberry according to claim 1, it is characterised in that: the formula of three kinds of culture mediums It is respectively as follows:
Initial culture base includes: MS culture medium, 0.5mg/L 6BA;
Subculture medium includes: MS culture medium, 0.5mg/L 6BA;
Root media includes: MS culture medium, 0.1mg/L IBA.
3. the method for tissue cultivating strawberry according to claim 1, it is characterised in that: in step S1, the ethyl alcohol is 75% Medical ethanol.
4. the method for tissue cultivating strawberry according to claim 1, it is characterised in that: in step S1, the mercury chloride is molten The concentration of liquid is 0.1%.
5. the method for tissue cultivating strawberry according to claim 1, it is characterised in that: in step S1, the sterile water For ultrapure water.
6. the method for tissue cultivating strawberry according to claim 1, it is characterised in that: it is 25 DEG C in temperature in step S2, Initial culture is carried out under dark conditions, incubation time is 30~35d;It is 25 DEG C in temperature, takes root under day light irradiation time 12h Culture, incubation time are 30~35d.
7. the method for tissue cultivating strawberry according to claim 1, it is characterised in that: in step S3, the sterilization matrix For coco bran: perlite=2:1 mixture.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113973718A (en) * 2021-12-22 2022-01-28 宿迁市设施园艺研究院 Strawberry tissue culture method
CN115517171A (en) * 2022-10-18 2022-12-27 太原师范学院 Method for producing strawberry seedlings by rapid plant tissue culture
CN116019012A (en) * 2022-12-29 2023-04-28 宿迁市设施园艺研究院 Detoxification seedling method for strawberries of snow rabbits
CN117305217A (en) * 2023-10-24 2023-12-29 临沂市农业科学院 Optimized stem tip tissue culture medium, culture method and application

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