CN109430062A - A kind of tissue culture method that large cherry is quickly bred - Google Patents
A kind of tissue culture method that large cherry is quickly bred Download PDFInfo
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- CN109430062A CN109430062A CN201811639735.3A CN201811639735A CN109430062A CN 109430062 A CN109430062 A CN 109430062A CN 201811639735 A CN201811639735 A CN 201811639735A CN 109430062 A CN109430062 A CN 109430062A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention belongs to large cherry tissue culture method fields, and in particular to a kind of tissue culture method that large cherry is quickly bred.The following steps are included: (1) takes required cherry rootstock treelet terminal bud, it is inoculated with after disinfection and cultivates on the induction medium；(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；(3) seedling in step (2) is cut, is inoculated in root media and cultivates；(4) it is cultivated on root media 10 days when seedling, when root long reaches 1-1.5cm, it takes out seedling and washes away root culture medium, move in Nutrition Soil, sprayed with 0.1% carbendazim saturating, it is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid removes plastic greenhouse after seven days when blade starts to grow, ventilation increases illumination, moves into crop field.The large cherry that this method is cultivated can not only fast-growth, the survival rate of large cherry can also be improved, in addition the sugar in nutrient solution also will increase the eating mouth feel of large cherry.
Description
Technical field
The invention belongs to large cherry tissue culture method fields, and in particular to a kind of tissue culture method that large cherry is quickly bred.
Background technique
Large cherry is a kind of rosaceae, cherry, and northern China deciduous fruit tree relays fruit maturation after cherry
Earliest fruit tree species.Traditional Chinese medicine and pharmacy thinks that large cherry has tonifying Qi in tune, the function of wind-damp dispelling.
The tissue culture technique of higher plant refers to that a part for separating one or several body cells or plant is trained
Feeding technology.The tissue cultures of broad sense usually described in us refer to a part by sterile working separating plant body, inoculation
It onto culture medium, is cultivated in the condition of manual control, it is made to generate complete plant.Tissue cultures can be divided by Object of Development
For tissue or callus culture, organ culture, plant culture, cell and Protoplast cuhnre etc..Wherein, plant culture is to complete
The culture of plant material, such as the culture of seedling and larger plant.
Quickly breed with the method for Plant Tissue Breeding is to produce upper most potential application, including flower ornamental is planted
Object, vegetables, fruit tree, field crop and other industrial crops.Fast breeding technique is not limited by conditions such as seasons, and growth cycle is short,
And the plant that cannot or be difficult breeding can be made to be proliferated.Quickly breeding can be carried out with following means: by stem apex, stem section,
The a large amount of axillary buds of the generations such as plateau；It is directly induced by organs such as root, leaves and generates adventitious bud；It is induced and is produced by callus tissue culture
Raw adventitious bud.Test tube rapid propagation is applied in following production or research: (1) breeding and hybridize on a small quantity obtained in crossbreeding
Kind, and save self-mating system, sterile line etc..(2) a small amount of virus-free seedling that breeding virus-free culture obtains.(3) breeding production is upper anxious
Seedling need or that provenance is less.Since the tissue cultures period is short, proliferation rate it is high and can whole year production the features such as, plus cultivating material
The miniaturization of material and test tube seedling, this can make limited space turn out a large amount of plant, turn out a large amount of children in a short time
Seedling.The prominent advantage of tissue cultures is " fast ", expands rapidly the quantity of plant in short period of time by this method.
Application No. is CN201510565330.X, a kind of large cherry is disclosed in the patent that publication date is 2015.11.11
Control root control be preced with cultural method, be related to fruit tree cultivation technical field.This method includes the steps that following sequences: (1) selection of land: selection
Plant place of the height above sea level in the cool area of the plateau 1800-2400m temperature；(2) seedling selects；(3) control root field planting；(4) fixed for the first time dry；
(5) first layer fruit-bearing shoot cluster culture；(6) it does calmly for second；(7) second layer fruit-bearing shoot cluster culture；(8) third time is fixed dry；(9)
Three-layered node fruit branch group culture；(10) the 4th times fixed dry；(11) the 4th layers of fruit-bearing shoot cluster culture；(12) it shapes.The present invention has can
With regulate and control root system, make lateral root shape is thick and short, tree crown photosynthesis sufficiently, fruit-bearing shoot cluster incubation time shorten 1-2 and also kind
Density in planting height, can significantly improve the beneficial effects such as cherry yield at Economization on land area.Although the invention have the advantages that it is above,
It is the problems such as there are still slow growths in the tissue culture procedures of large cherry, therefore, it is necessary to improve to the program.
Summary of the invention
Aiming at the shortcomings in the prior art, the present invention provides a kind of tissue culture method that large cherry is quickly bred, in this method
By taking required cherry rootstock treelet terminal bud, it is successively seeded in the induced medium containing different component, Multiplying culture
On base and root media, finally apply nutrient solution again so that large cherry can not only fast-growth, large cherry can also be improved
Survival rate, in addition the sugar in nutrient solution also will increase the eating mouth feel of large cherry.
In order to achieve the above-mentioned object of the invention, the invention adopts the following technical scheme:
A kind of tissue culture method that large cherry is quickly bred, comprising the following steps:
(1) required cherry rootstock treelet terminal bud is taken, is inoculated with after disinfection and cultivates on the induction medium；
(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；
(3) seedling in step (2) is cut, is inoculated in root media and cultivates；
(4) it is cultivated on root media 10 days, when root long reaches 1-1.5cm when seedling, take out seedling and washes away root
Culture medium moves in Nutrition Soil, is sprayed thoroughly, is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid works as leaf with 0.1% carbendazim
Piece starts to grow, and plastic greenhouse is removed after seven days, and ventilation increases illumination, moves into crop field.
Preferably, step (1) the Fiber differentiation based formulas and each raw material proportioning are as follows: be added in 1 liter of 1/2MS culture medium
Following raw materials and its proportion are made: 0.5~0.9mg of agar, 0.2~0.5mg of white granulated sugar, 6- benzyl aminoadenine 0.02~
0.05mg, 0.01~0.03mg of methyl α-naphthyl acetate, 0.05~0.09mg of carbenicillin, 10~15mg of coconut milk, active carbon 0.8~
1.2mg。
Preferably, step (1) condition of culture: temperature is 23 ± 1 DEG C, and light intensity is 2000~2500Lx, the time 18 is small
When/day.
Preferably, step (2) proliferation culture medium formula and each raw material proportioning are as follows: under being added in 1 liter of MS culture medium
It states raw material and its proportion is made: 0.6~0.8mg of carragheen, 3.2~5.0mg of sucrose, inositol 0.02~0.06mg and NAA
0.015mg。
Preferably, step (2) condition of culture: 25 ± 2 DEG C of temperature, 2500~3000Lx of light intensity, 16 hours time/
It.
Preferably, step (3) prescription of rooting medium and each raw material proportioning are as follows: minimal medium 1/3MS, indoles fourth
Sour 0.9-1.2mg, 1- methyl α-naphthyl acetate 0.05-0.1mg, sucrose 30-45mg, agar powder 45-50mg.
Preferably, step (4) Nutrition Soil is the mixture formed containing vermiculite matrix, fine sand, decomposed manure.
Preferably, ammonium nitrate 1650mg, potassium nitrate 1900mg, potassium dihydrogen phosphate are contained in step (4) nutrient solution
170mg, magnesium sulfate 370mg, calcium chloride 440mg, potassium iodide 0.41mg, boric acid 3.1mg, manganese sulfate 11.1mg, zinc sulfate
4.3mg, sodium molybdate 0.125mg, copper sulphate 0.0125mg, cobalt chloride 0.0125mg, indolebutyric acid 0.5mg, heteroauxin
0.4mg。
Compared with prior art, the invention has the following advantages:
By taking required large cherry stock treelet terminal bud in the program, it is successively seeded in luring containing different component
Lead on culture medium, proliferated culture medium and root media, finally apply nutrient solution again so that large cherry can not only fast-growth,
The survival rate of large cherry can also be improved, in addition the sugar in nutrient solution also will increase the eating mouth feel of large cherry；Fiber differentiation
The each component of base cooperates, and the survival rate of large cherry not only can be improved, and have it is at low cost, explant after sterilizing can be effectively improved
The characteristics of body survival rate；Proliferated culture medium significantly shortens the Multiplying culture time, improves Multiplying culture efficiency, root media
Improvement so that the rootage duration of large cherry foreshortened to 10 days from 15 days, Nutrition Soil be large cherry seedling root growth create compared with
Good condition, the nutrient solution in addition containing various plants indispensable element promote the growth of blade.
Specific embodiment
Below by specific embodiment, invention is further described in detail.But those skilled in the art will manage
Solution, the following example is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Specific skill is not specified in embodiment
Art or condition person, described technology or conditions carry out to specifications according to the literature in the art.Agents useful for same or instrument
Production firm person is not specified, being can be with conventional products that are commercially available.
Embodiment 1
A kind of tissue culture method that large cherry is quickly bred, comprising the following steps:
(1) required cherry rootstock treelet terminal bud is taken, is inoculated with after disinfection and cultivates on the induction medium；
(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；
(3) seedling in step (2) is cut, is inoculated in root media and cultivates；
(4) it is cultivated on root media 10 days, when root long reaches 1cm when seedling, take out seedling and washes away root culture
Base moves in Nutrition Soil, is sprayed thoroughly, is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid, when blade is opened with 0.1% carbendazim
Begin to grow, plastic greenhouse is removed after seven days, ventilation increases illumination, moves into crop field.
Specifically, Fiber differentiation based formulas and each raw material proportioning are as follows: be added in 1 liter of 1/2MS culture medium following raw materials and
Its proportion is made: agar 0.5mg, white granulated sugar 0.5mg, 6- benzyl aminoadenine 0.02mg, methyl α-naphthyl acetate 0.03mg, carbenicillin
0.05mg, coconut milk 15mg, active carbon 0.8mg；Temperature is 24 DEG C in condition of culture, light intensity 2500Lx, 18 hours time/
It.
Proliferation culture medium formula and each raw material proportioning are as follows: following raw materials are added in 1 liter of MS culture medium and its proportion is made:
Carragheen 0.6mg, sucrose 3.8mg, inositol 0.03mg and NAA 0.015mg, condition of culture: 25 DEG C of temperature, light intensity 2500Lx, when
Between 16 hours/day.
Prescription of rooting medium and each raw material proportioning are as follows: minimal medium 1/3MS, indolebutyric acid 0.9mg, 1- methyl α-naphthyl acetate
0.09mg, sucrose 35mg, agar powder 50mg.
Nutrition Soil is the mixture formed containing vermiculite matrix, fine sand, decomposed manure.
Contain ammonium nitrate 1650mg, potassium nitrate 1900mg, potassium dihydrogen phosphate 170mg, magnesium sulfate 370mg, chlorine in nutrient solution
Change calcium 440mg, potassium iodide 0.41mg, boric acid 3.1mg, manganese sulfate 11.1mg, zinc sulfate 4.3mg, sodium molybdate 0.125mg, sulfuric acid
Copper 0.0125mg, cobalt chloride 0.0125mg, indolebutyric acid 0.5mg, heteroauxin 0.4mg.
Embodiment 2
A kind of tissue culture method that large cherry is quickly bred, comprising the following steps:
(1) required cherry rootstock treelet terminal bud is taken, is inoculated with after disinfection and cultivates on the induction medium；
(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；
(3) seedling in step (2) is cut, is inoculated in root media and cultivates；
(4) it is cultivated on root media 10 days, when root long reaches 1cm when seedling, take out seedling and washes away root culture
Base moves in Nutrition Soil, is sprayed thoroughly, is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid, when blade is opened with 0.1% carbendazim
Begin to grow, plastic greenhouse is removed after seven days, ventilation increases illumination, moves into crop field.
Specifically, Fiber differentiation based formulas and each raw material proportioning are as follows: be added in 1 liter of 1/2MS culture medium following raw materials and
Its proportion is made: agar 0.6mg, white granulated sugar 0.3mg, 6- benzyl aminoadenine 0.03mg, methyl α-naphthyl acetate 0.025mg, carboxylic benzyl mould
Plain 0.06mg, coconut milk 13mg, active carbon 0.9mg；Temperature is 22 DEG C in condition of culture, light intensity 2200Lx, 18 hours time/
It.
Proliferation culture medium formula and each raw material proportioning are as follows: following raw materials are added in 1 liter of MS culture medium and its proportion is made:
Carragheen 0.7mg, sucrose 3.5mg, inositol 0.04mg and NAA 0.015mg, condition of culture: 26 DEG C of temperature, light intensity 2800Lx, when
Between 16 hours/day.
Prescription of rooting medium and each raw material proportioning are as follows: minimal medium 1/3MS, indolebutyric acid 1.0mg, 1- methyl α-naphthyl acetate
0.05mg, sucrose 40mg, agar powder 48mg.
Nutrition Soil is the mixture formed containing vermiculite matrix, fine sand, decomposed manure.
Contain ammonium nitrate 1650mg, potassium nitrate 1900mg, potassium dihydrogen phosphate 170mg, magnesium sulfate 370mg, chlorine in nutrient solution
Change calcium 440mg, potassium iodide 0.41mg, boric acid 3.1mg, manganese sulfate 11.1mg, zinc sulfate 4.3mg, sodium molybdate 0.125mg, sulfuric acid
Copper 0.0125mg, cobalt chloride 0.0125mg, indolebutyric acid 0.5mg, heteroauxin 0.4mg.
Embodiment 3
A kind of tissue culture method that large cherry is quickly bred, comprising the following steps:
(1) required cherry rootstock treelet terminal bud is taken, is inoculated with after disinfection and cultivates on the induction medium；
(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；
(3) seedling in step (2) is cut, is inoculated in root media and cultivates；
(4) it is cultivated on root media 10 days, when root long reaches 1cm when seedling, take out seedling and washes away root culture
Base moves in Nutrition Soil, is sprayed thoroughly, is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid, when blade is opened with 0.1% carbendazim
Begin to grow, plastic greenhouse is removed after seven days, ventilation increases illumination, moves into crop field.
Specifically, Fiber differentiation based formulas and each raw material proportioning are as follows: be added in 1 liter of 1/2MS culture medium following raw materials and
Its proportion is made: agar 0.7mg, white granulated sugar 0.4mg, 6- benzyl aminoadenine 0.04mg, methyl α-naphthyl acetate 0.01mg, carbenicillin
0.07mg, coconut milk 12mg, active carbon 1.1mg；Temperature is 24 DEG C in condition of culture, light intensity 2200Lx, 18 hours time/
It.
Proliferation culture medium formula and each raw material proportioning are as follows: following raw materials are added in 1 liter of MS culture medium and its proportion is made:
Carragheen 0.75mg, sucrose 4.5mg, inositol 0.05mg and NAA 0.015mg, condition of culture: 24 DEG C of temperature, light intensity 2600Lx,
16 hour/day of time.
Prescription of rooting medium and each raw material proportioning are as follows: minimal medium 1/3MS, indolebutyric acid 1.1mg, 1- methyl α-naphthyl acetate
0.08mg, sucrose 45mg, agar powder 45mg.
Nutrition Soil is the mixture formed containing vermiculite matrix, fine sand, decomposed manure.
Contain ammonium nitrate 1650mg, potassium nitrate 1900mg, potassium dihydrogen phosphate 170mg, magnesium sulfate 370mg, chlorine in nutrient solution
Change calcium 440mg, potassium iodide 0.41mg, boric acid 3.1mg, manganese sulfate 11.1mg, zinc sulfate 4.3mg, sodium molybdate 0.125mg, sulfuric acid
Copper 0.0125mg, cobalt chloride 0.0125mg, indolebutyric acid 0.5mg, heteroauxin 0.4mg.
Embodiment 4
A kind of tissue culture method that large cherry is quickly bred, comprising the following steps:
(1) required cherry rootstock treelet terminal bud is taken, is inoculated with after disinfection and cultivates on the induction medium；
(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；
(3) seedling in step (2) is cut, is inoculated in root media and cultivates；
(4) it is cultivated on root media 10 days, when root long reaches 1cm when seedling, take out seedling and washes away root culture
Base moves in Nutrition Soil, is sprayed thoroughly, is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid, when blade is opened with 0.1% carbendazim
Begin to grow, plastic greenhouse is removed after seven days, ventilation increases illumination, moves into crop field.
Specifically, Fiber differentiation based formulas and each raw material proportioning are as follows: be added in 1 liter of 1/2MS culture medium following raw materials and
Its proportion is made: agar 0.9mg, white granulated sugar 0.2mg, 6- benzyl aminoadenine 0.05mg, methyl α-naphthyl acetate 0.01mg, carbenicillin
0.09mg, coconut milk 10mg, active carbon 1.2mg；Temperature is 24 DEG C in condition of culture, light intensity 2500Lx, 18 hours time/
It.
Proliferation culture medium formula and each raw material proportioning are as follows: following raw materials are added in 1 liter of MS culture medium and its proportion is made:
Carragheen 0.8mg, sucrose 3.2mg, inositol 0.06mg and NAA 0.015mg, condition of culture: 27 DEG C of temperature, light intensity 2900Lx, when
Between 16 hours/day.
Prescription of rooting medium and each raw material proportioning are as follows: minimal medium 1/3MS, indolebutyric acid 0.9mg, 1- methyl α-naphthyl acetate
0.09mg, sucrose 45mg, agar powder 47mg.
Nutrition Soil is the mixture formed containing vermiculite matrix, fine sand, decomposed manure.
Contain ammonium nitrate 1650mg, potassium nitrate 1900mg, potassium dihydrogen phosphate 170mg, magnesium sulfate 370mg, chlorine in nutrient solution
Change calcium 440mg, potassium iodide 0.41mg, boric acid 3.1mg, manganese sulfate 11.1mg, zinc sulfate 4.3mg, sodium molybdate 0.125mg, sulfuric acid
Copper 0.0125mg, cobalt chloride 0.0125mg, indolebutyric acid 0.5mg, heteroauxin 0.4mg.
The above is only preferred embodiments of the present invention, is not intended to limit the scope of the present invention,
Therefore any trickle amendment, equivalent variations and modification made to the above embodiment according to the technical essence of the invention, belong to
In the range of technical solution of the present invention.
Claims (8)
1. a kind of tissue culture method that large cherry is quickly bred, which comprises the following steps:
(1) required cherry rootstock treelet terminal bud is taken, is inoculated with after disinfection and cultivates on the induction medium；
(2) it takes and grows bud clump stem apex in step (1), be inoculated on proliferated culture medium and cultivate；
(3) seedling in step (2) is cut, is inoculated in root media and cultivates；
(4) it is cultivated on root media 10 days, when root long reaches 1-1.5cm when seedling, take out seedling and washes away root culture
Base moves in Nutrition Soil, is sprayed thoroughly, is placed in plastic greenhouse, every 2 days sprinkling one time of nutrition liquid, when blade is opened with 0.1% carbendazim
Begin to grow, plastic greenhouse is removed after seven days, ventilation increases illumination, moves into crop field.
2. the tissue culture method that a kind of large cherry according to claim 1 is quickly bred, which is characterized in that the step (1)
The formula of induced medium and each raw material proportioning are as follows: following raw materials are added in 1 liter of 1/2MS culture medium and its proportion is made: fine jade
0.5~0.9mg of rouge, 0.2~0.5mg of white granulated sugar, 0.02~0.05mg of 6- benzyl aminoadenine, 0.01~0.03mg of methyl α-naphthyl acetate,
0.05~0.09mg of carbenicillin, 10~15mg of coconut milk, 0.8~1.2mg of active carbon.
3. the tissue culture method that a kind of large cherry according to claim 1 or 2 is quickly bred, which is characterized in that the step
(1) condition of culture: temperature is 23 ± 1 DEG C, and light intensity is 2000~2500Lx, 18 hour/day of time.
4. the tissue culture method that a kind of large cherry according to claim 1 is quickly bred, which is characterized in that the step (2)
Proliferation culture medium formula and each raw material proportioning are as follows: following raw materials are added in 1 liter of MS culture medium and its proportion is made: carragheen
0.6~0.8mg, 3.2~5.0mg of sucrose, inositol 0.02~0.06mg and NAA 0.015mg.
5. the tissue culture method that a kind of large cherry according to claim 1 or 4 is quickly bred, which is characterized in that the step
(2) condition of culture: 25 ± 2 DEG C of temperature, 2500~3000Lx of light intensity, 16 hour/day of time.
6. the tissue culture method that a kind of large cherry according to claim 1 is quickly bred, which is characterized in that the step (3)
Prescription of rooting medium and each raw material proportioning are as follows: minimal medium 1/3MS, indolebutyric acid 0.9-1.2mg, 1- methyl α-naphthyl acetate 0.05-
0.1mg, sucrose 30-45mg, agar powder 45-50mg.
7. the tissue culture method that a kind of large cherry according to claim 1 is quickly bred, which is characterized in that the step (4)
Nutrition Soil is the mixture formed containing vermiculite matrix, fine sand, decomposed manure.
8. the tissue culture method that a kind of large cherry according to claim 1 is quickly bred, which is characterized in that the step (4)
Contain ammonium nitrate 1650mg, potassium nitrate 1900mg, potassium dihydrogen phosphate 170mg, magnesium sulfate 370mg, calcium chloride in nutrient solution
440mg, potassium iodide 0.41mg, boric acid 3.1mg, manganese sulfate 11.1mg, zinc sulfate 4.3mg, sodium molybdate 0.125mg, copper sulphate
0.0125mg, cobalt chloride 0.0125mg, indolebutyric acid 0.5mg, heteroauxin 0.4mg.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111011211A (en) * | 2019-12-26 | 2020-04-17 | 南京千亩休闲农业发展有限公司 | Novel sweet cherry insect-resistant fruit tree cultivation method |
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Application publication date: 20190308 |