CN105941158B - A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology - Google Patents
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology Download PDFInfo
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Classifications
-
- 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
Abstract
The present invention discloses a kind of method that strawberry detoxic seedling is cultivated using tissue culture technology, including：(1) explant is chosen and sterilized：The unopened bud of strawberrying plant, after alcohol and mercuric chloride sterilization treatment, flower pesticide is taken out, obtains explant；(2) inducing clumping bud：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 8-10 days, then illumination cultivation 80-90 days, until produce Multiple Buds, and the bud of Multiple Buds high is not less than 2cm；(3) Multiple Buds switching culture：Multiple Buds obtained by step (2) are forwarded in culture medium M2, cultivated 15-25 days, until Multiple Buds grow up to the high Strawberry seedlings of 3-5cm, then the Strawberry seedlings are forwarded in culture medium M3, culture of rootage is carried out, produces strawberry detoxic seedling.Used medium formula of the present invention is simple, and tissue culture simple flow, virus elimination rate reaches 100%, breeds coefficient height, transplanting survival rate is high, and seedling is neat, is easy to transplanting and Cultivate administration that the later stage is unified, can carry out industrialization production.
Description
Technical field
The present invention relates to biological tissue culture technical field, and in particular to one kind cultivates strawberry detoxic seedling using tissue culture technology
Method.
Background technology
Strawberry is rose family Fragaria perennial root perennial evergreen herbaceous plant, belongs to polyploid, is that one kind leans on stolon
The crop of vegetative propagation is carried out, the method for traditional seedling fostering is to use the vegetative manner for stem breeding of crawling, less efficient,
It is unfavorable for the popularization of improved seeds, and easily virus infection.Identify that clear and definite strawberry virus has four kinds, i.e. strawberry spot in China
Refute viral (SMOV), strawberry light yellow edge virus (SMYEV), strawberry crinkle virus (SCRV) and strawberry veinbanding virus (SVBV).
The universal virus infection of seedling is the bottleneck problem in strawberry production, the strawberry growing way of the strawberry of virus infection than uninfecting virus
It is weak, resistance is poor, yield is decreased obviously, nutrient quality deteriorates with commodity property.For virosis, there is presently no medicament to control
Reason, therefore, cultivates virus-free female parent seedling, cultivates virus-free nursery stock, is the fundamental measure for preventing and treating Strawberry Virus.At present, it is disease-free
Malicious female parent seedling mainly passes through strawberry detoxification tissue culture technology, i.e., is taken off by technologies such as anther tissue culture, Micro-stem tip tissue cultures
Virus removal cause of disease obtains nontoxic strawberry regeneration plant, strawberry seedling is recovered kind original seed good strains of seeds, to reach high-quality, high yield
Purpose.But the virus-free female parent seedling of strawberry is cultivated and is not easy, the explant of tissue cultures selects defective, the bar in incubation
Part control deficiency etc., it will result in poor detoxification efficiency, pollution, vitrifying, breed the problems such as coefficient is low, transplanting survival rate is not high,
Very big loss is caused to production.
The content of the invention
In view of this, the application provides a kind of method that strawberry detoxic seedling is cultivated using tissue culture technology, and used medium is matched somebody with somebody
Side is simple, and tissue culture simple flow, virus elimination rate reaches 100%, breeds coefficient height, transplanting survival rate is high, and seedling is neat, is easy to the later stage
Unified transplanting and Cultivate administration, reduce cost of labor, can carry out industrialization production.
To solve above technical problem, technical scheme provided by the invention is that one kind cultivates strawberry detoxification using tissue culture technology
The method of seedling, the described method comprises the following steps：
(1) explant is chosen and sterilized：The unopened bud of strawberrying plant, using alcohol and mercuric chloride sterilization treatment
Afterwards, bud is peeled off, flower pesticide is taken out, obtains explant；
(2) inducing clumping bud：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 8-10 days, then
Illumination cultivation 80-90 days, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm；The composition of the culture medium M1 is：
MS minimal mediums, supplement 0.5-3.0mg/L TDZ, 0.05-0.30mg/L IAA；
(3) Multiple Buds switching culture：Multiple Buds obtained by step (2) are forwarded in culture medium M2, culture 15-25 days, directly
Grow up to the high Strawberry seedlings of 3-5cm to Multiple Buds, then the Strawberry seedlings be forwarded in culture medium M3, carry out culture of rootage,
Produce strawberry detoxic seedling；The culture medium M2 is MS minimal mediums, and the culture medium M3 is 1/2MS minimal mediums.
Preferably, in step (1), the sterilising conditions of the bud are：First sterilized 18-25s, then used with 75% alcohol
0.1% mercuric chloride solution sterilizing, 12-15min.
It is more highly preferred to, in step (1), the sterilising conditions of the flower pesticide are：First sterilized 20s, then used with 75% alcohol
0.1% mercuric chloride solution sterilizing 13min.
Preferably, in step (2), the explant is inoculated in culture medium M1, first light culture 9 days, then illumination cultivation 80
My god.
Preferably, in step (2), the composition of the culture medium M1 is：MS minimal mediums, supplement 1.0-2.0mg/L
TDZ、0.1—0.2mg/L IAA。
Preferably, in step (2), the condition of culture is 2000-3000LX of intensity of illumination, light irradiation time 16h/ days, is trained
Support 24-26 DEG C of temperature.
Preferably, in step (3), the Multiple Buds are forwarded in culture medium M2, are cultivated 20 days.
Preferably, in step (3), the Multiple Buds are in culture medium M2 or the Strawberry seedlings are in culture medium M3 training
Foster condition is：2000-3000LX of intensity of illumination, light irradiation time 16h/ days, 24-26 DEG C of cultivation temperature.
Preferably, in step (1), the kind of the strawberry is selected from a liquor-saturated chivalrous, snow younger sister, chapter Ji, more beautiful for any one.
In technical scheme, the TDZ is a plant growth regulators, and there is the very strong basic element of cell division to live
Property, the regeneration and breeding of plant sprout can be promoted, break the eye of stopping of bud, promote seed to sprout, promote callus growth, delayed
Plant senescence etc., the growth and development process of plant can be adjusted to the effect of other plant hormones and physiological activator,
It can be used as Plant Tissue Breeding.
The IAA is heteroauxin, is a kind of auxin, can adjust the growth of plant, can not only promote to give birth to
It is long, and have the function that to suppress growth and Apparatuses formation.On a cellular level, cambial cell division can be stimulated, stimulate branch
Cell elongation, suppress root cell growth, promote xylem, phloem cell differentiation, promote cutting root of hair, regulation callus
Morphogenesis, therefore can be used as Plant Tissue Breeding.
The MS minimal mediums have higher inorganic salt concentration, can ensure the mineral nutrition needed for tissue growth,
The growth of callus can also be accelerated, be more stable ionic equilibrium solution, its nitrate content is high, the quantity of its nutrient and
Ratio is suitable, can meet the nutrition and physiological requirements of plant cell, thus the scope of application is wider, most plants tissue culture rapid
Speed breeding uses it as the minimal medium of culture medium.
The 1/2MS minimal mediums are that a great number of elements halves, the culture of remaining constant formation in MS minimal mediums
Base.
Technical scheme provides a kind of method that strawberry detoxic seedling is cultivated using tissue culture technology, including explant choosing
Take and sterilize, inducing clumping bud, Multiple Buds switching culture the step of, using flower pesticide as explant progress tissue cultures, by flower
Medicine is inoculated in culture medium M1 when being cultivated, and first carries out light culture, then carries out illumination cultivation, and illumination cultivation was opened to 10 days or so
There is callus in beginning, and illumination cultivation was to 40 days or so, callus length to 1cm or so, and illumination cultivation was to 60 days or so, callus
Tissue is long to diameter 2-3cm sizes, and has small bud point to sprout, and for illumination cultivation to 80 days or so, small bud point grew blade, and bud is high
2cm or so, the bud grown is forwarded in culture medium M2, until bud grows up to the high Strawberry seedlings of 3-5cm, then is forwarded to culture
Base M3 carries out culture of rootage, produces complete strawberry detoxification tissue culture seedling.
Snow younger sister's kind described herein, fruit shape is good, color is beautiful, sugariness is high, manageability, compared with planting traditional strawberry,
It is small to dredge leaf amount, without flower thinning, vegetables and fruits, artificial 20% can be saved；Disease resistance is strong, malformed fruit rate is low, can increase production 20%.
Described chapter Ji's kind, fruit are in neatly conico-acuminate, and fruit is healthy and strong, bright in colour bright, good smell；Pulp
Pale red, delicate succulence, dense sweet taste, enjoy endless aftertastes, the superfine product being described as in Japan in strawberry；Bear fruit strong, it is ripe early, do not resist
Anthrax.
Described liquor-saturated chivalrous kind, plant strain growth gesture is strong, and stolon generating capacity is few, and fruit is peony, and fruit shape is good；Yielding ability
Good, fruit is in conico-acuminate, and fruit shape is big, no ditch, no malformed fruit, fruit ruby, good luster, pulp white, the crisp and refreshing mouth of sweet tea；Height is anti-
Gray mold, powdery mildew and anthracnose.
Described more beautiful kind, shallow dormancy, precocity, plant type are compacter；Early yield is high, commodity is good, quality with it is " red
Cheek " kind is close；Feel anthracnose, mildew-resistance.
Above-mentioned strawberry cultivars can use the cultivation of the method progress strawberry detoxic seedling described in technical scheme.
Compared to existing tissue culture technology, technical scheme used medium formula is simple, tissue culture simple flow, detoxification
Rate reaches 100%, breeds coefficient height, and transplanting survival rate is high, and seedling is neat, is easy to transplanting and Cultivate administration that the later stage is unified, drop
Low cost of labor, can carry out industrialization production.
Embodiment
In order that those skilled in the art more fully understands technical scheme, with reference to specific embodiment pair
The present invention is described in further detail.
Embodiment 1
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, is comprised the following steps：
(1) explant is chosen and sterilized：Gather unopened bud of the kind for the strawberry of snow younger sister, the size of bud
In 0.4-0.6mm, first sterilized 20s with 75% alcohol, then sterilized 13min with 0.1% mercuric chloride solution, taken out flower pesticide, obtain explant
Body；
(2) inducing clumping bud：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 9 days, then illumination
Culture 80 days, until produce Multiple Buds, and Multiple Buds bud it is high be not less than 2cm, condition of culture for cultivate illumination 2000-
3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The composition of the culture medium M1 is：MS minimal mediums, supplement
1.0mg/L TDZ、0.2mg/L IAA；
(3) Multiple Buds switching culture：Multiple Buds obtained by step (2) are forwarded in culture medium M2, cultivated 20 days, until clump
Sprout and grow up to the high Strawberry seedlings of 3-5cm, then the Strawberry seedlings are forwarded in culture medium M3, carry out culture of rootage, produce
Strawberry detoxic seedling；The culture medium M2 is MS minimal mediums, and condition of culture is to cultivate 2000-3000LX of illumination, cultivation temperature
24-26 DEG C, light irradiation time 16h/ days, the culture medium M3 is 1/2MS minimal mediums, and condition of culture is culture illumination
2000-3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days.
The present embodiment methods described cultivates strawberry detoxic seedling, and time of its callus generation is light culture light again after 9 days
According to culture 10 days, illumination cultivation occurred bud point, 80 days or so bud point length of culture to 2cm or so for 60 days or so, and callus rate is
71.1%, differentiation rate is 52.3%, virus elimination rate 100%, breeding coefficient 7, survival rate of plant 92%.
Embodiment 2
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (2) inducing clumping bud is：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 9 days,
Illumination cultivation 80 days again, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm, the condition of culture is culture illumination
2000-3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The composition of the culture medium M1 is：MS is cultivated substantially
Base, supplement 1.5mg/L TDZ, 0.1mg/L IAA.
The present embodiment methods described cultivates strawberry detoxic seedling, and explant is inoculated in when being cultivated in culture medium M1, callus
The time of tissue generation, illumination cultivation 10 days, illumination cultivation occurred bud point for 60 days or so again after 9 days for light culture, cultivated 80 days left sides
Right bud point length is to 2cm or so, and callus rate is 71.1%, and differentiation rate is 52.3%, virus elimination rate 100%, breeding coefficient
For 7, survival rate of plant 92%.
Embodiment 3
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (2) inducing clumping bud is：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 9 days,
Illumination cultivation 80 days again, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm, the condition of culture is culture illumination
2000-3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The composition of the culture medium M1 is：MS is cultivated substantially
Base, supplement 1.0mg/L TDZ, 0.2mg/L IAA.
The present embodiment methods described cultivates strawberry detoxic seedling, and explant is inoculated in when being cultivated in culture medium M1, callus
The time of tissue generation, illumination cultivation 10 days, illumination cultivation occurred bud point for 60 days or so again after 9 days for light culture, cultivated 80 days left sides
Right bud point length is to 2cm or so, and callus rate is 79.2%, and differentiation rate is 55.6%, virus elimination rate 100%, breeding coefficient
For 8, survival rate of plant 95.6%.
Embodiment 4
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (2) inducing clumping bud is：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 9 days,
Illumination cultivation 80 days again, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm, the condition of culture is culture illumination
2000-3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The composition of the culture medium M1 is：MS is cultivated substantially
Base, supplement 1.5mg/L TDZ, 0.1mg/L IAA.
The present embodiment methods described cultivates strawberry detoxic seedling, and explant is inoculated in when being cultivated in culture medium M1, callus
The time of tissue generation, illumination cultivation 10 days, illumination cultivation occurred bud point for 60 days or so again after 9 days for light culture, cultivated 80 days left sides
Right bud point length is to 2cm or so, and callus rate is 79.2%, and differentiation rate is 55.6%, virus elimination rate 100%, breeding coefficient
For 8, survival rate of plant 95.6%.
Embodiment 5
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (2) inducing clumping bud is：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 9 days,
Illumination cultivation 80 days again, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm, the condition of culture is culture illumination
2000-3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The composition of the culture medium M1 is：MS is cultivated substantially
Base, supplement 1.5mg/L TDZ, 0.2mg/L IAA.
The present embodiment methods described cultivates strawberry detoxic seedling, and explant is inoculated in when being cultivated in culture medium M1, callus
The time of tissue generation, illumination cultivation 10 days, illumination cultivation occurred bud point for 60 days or so again after 9 days for light culture, cultivated 80 days left sides
Right bud point length is to 2cm or so, and callus rate is 82.2%, and differentiation rate is 59.8%, virus elimination rate 100%, breeding coefficient
For 7, survival rate of plant 96.7%.
Embodiment 6
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (2) inducing clumping bud is：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 9 days,
Illumination cultivation 80 days again, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm, the condition of culture is culture illumination
2000-3000LX, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The composition of the culture medium M1 is：MS is cultivated substantially
Base, supplement 0.5mg/L TDZ, 0.10mg/L IAA.
The present embodiment methods described cultivates strawberry detoxic seedling, and explant is inoculated in when being cultivated in culture medium M1, callus
The time of tissue generation, illumination cultivation 10 days, illumination cultivation occurred bud point for 60 days or so again after 9 days for light culture, cultivated 80 days left sides
Right bud point length is to 2cm or so, and callus rate is 83.6%, and differentiation rate is 60.5%, virus elimination rate 100%, breeding coefficient
For 7, survival rate of plant 96.8%.
Embodiment 7
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (1) explant is chosen and sterilizing is：Gather the unopened bud that kind is liquor-saturated chivalrous strawberry, bud
Size in 0.4-0.6mm, sterilized 18s with 75% alcohol, then sterilized 12min with 0.1% mercuric chloride solution, take out flower pesticide, obtain outer
Implant.
The present embodiment methods described cultivates strawberry detoxic seedling, and the time of callus generation is light culture illumination again after 9 days
Culture 10 days, illumination cultivation occur bud point, 80 days or so bud point length of culture to 2cm or so for 60 days or so, and callus rate is 88.9%,
Differentiation rate is 61.1%, virus elimination rate 100%, breeding coefficient 8, survival rate of plant 100%.
Embodiment 8
A kind of method that strawberry detoxic seedling is cultivated using tissue culture technology described in the present embodiment, with the methods described of embodiment 1
Difference be：
Step (1) explant is chosen and sterilizing is：Gather the unopened bud that kind is more beautiful strawberry, bud
Size in 0.4-0.6mm, first sterilized 25s with 75% alcohol, then sterilized 15min with 0.1% mercuric chloride solution, take out flower pesticide, obtain
Explant.
The present embodiment methods described cultivates strawberry detoxic seedling, and the time of callus generation is light culture illumination again after 9 days
Culture 10 days, illumination cultivation occur bud point, 80 days or so bud point length of culture to 2cm or so for 60 days or so, and callus rate is 84.5%,
Differentiation rate is 60.2%, virus elimination rate 100%, breeding coefficient 7, survival rate of plant 97.2%.
Embodiment 9
The influence of growth hormone composition and concentration to cultivation in culture medium M1
Take the consistent explant of the growing state of different strawberry cultivars (snow younger sister, a more beautiful, liquor-saturated chivalrous, chapter Ji) some, connect respectively
Kind into culture medium M1, first light culture 9 days, then illumination cultivation 80 days, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than
2cm, the condition of culture are to cultivate 2000-3000LX of illumination, 24-26 DEG C of cultivation temperature, light irradiation time 16h/ days；The culture
Base M1 supplements certain growth hormone and formed using MS minimal mediums.In the MS minimal mediums identical condition
Under, culture medium numbering is carried out according to the composition of growth hormone and concentration, respectively A, B, C, D, E, F, G, specific numbering situation is shown in
Table 1, observe and record the culture situation of each strawberry cultivars explant in each culture medium, cultivation results are shown in Table 2-5.
The hormonal components and concentration of each culture medium in the embodiment 9 of table 1
| Culture medium is numbered | TDZ(mg/L) | IAA(mg/L) |
| A | -- | 0.15 |
| B | 1.5 | -- |
| C | 0.5 | 0.05 |
| D | 1.0 | 0.1 |
| E | 1.5 | 0.15 |
| F | 2.0 | 0.2 |
| G | 3.0 | 0.3 |
Table 2 avenges the younger sister's kind influence result of growth hormone composition and concentration to culture in each culture medium
| A | B | C | D | E | F | G | |
| Inoculation number () | 18 | 18 | 18 | 18 | 18 | 18 | 18 |
| Callus number (individual) | 3 | 4 | 8 | 11 | 14 | 12 | 9 |
| Callus rate (%) | 16.7 | 22.2 | 44.4 | 61.1 | 77.8 | 66.7 | 50.0 |
| Multiple Buds number (individual) | 0 | 0 | 1 | 5 | 11 | 4 | 3 |
| Differentiation rate (%) | 0.0 | 0.0 | 5.6 | 27.8 | 61.1 | 22.2 | 16.7 |
| Death toll () | 15 | 14 | 10 | 7 | 4 | 6 | 9 |
| The death rate (%) | 83.3 | 77.8 | 55.6 | 38.9 | 22.2 | 33.3 | 50.0 |
3 more beautiful kind of the table influence result of growth hormone composition and concentration to culture in each culture medium
The liquor-saturated chivalrous kind of the table 4 influence result of growth hormone composition and concentration to culture in each culture medium
| A | B | C | D | E | F | G | |
| Inoculation number () | 18 | 18 | 18 | 18 | 18 | 18 | 18 |
| Callus number (individual) | 1 | 2 | 4 | 10 | 16 | 11 | 6 |
| Callus rate (%) | 5.6 | 11.1 | 22.2 | 55.6 | 88.9 | 61.1 | 33.3 |
| Multiple Buds number (individual) | 0 | 0 | 1 | 5 | 11 | 4 | 0 |
| Differentiation rate (%) | 0.0 | 0.0 | 5.6 | 27.8 | 61.1 | 22.2 | 0.0 |
| Death toll () | 17 | 16 | 14 | 8 | 2 | 7 | 12 |
| The death rate (%) | 94.4 | 88.9 | 77.8 | 44.4 | 11.1 | 38.9 | 66.7 |
The chapter Ji kind of the table 5 influence result of growth hormone composition and concentration to culture in each culture medium
As can be seen from the above data, TDZ and IAA uses simultaneously are more preferable than alone one of which, its callus rate, life of growing thickly
Inductivity and survival rate of plant be intended to it is higher, wherein TDZ in 1.0-2.0mg/L, IAA in 0.1-0.2mg/L, particularly
For TDZ in 1.5mg/L, IAA its indices in 0.15mg/L are preferable, are the preferred scheme of the application.
It the above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
The limitation of the present invention, protection scope of the present invention should be defined by claim limited range.For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, some improvements and modifications can also be made, these change
Enter and retouch and also should be regarded as protection scope of the present invention.
Claims (8)
- A kind of 1. method that strawberry detoxic seedling is cultivated using tissue culture technology, it is characterised in that：It the described method comprises the following steps：(1) explant is chosen and sterilized：The unopened bud of strawberrying plant, after alcohol and mercuric chloride sterilization treatment, take out Flower pesticide, obtain explant；(2) inducing clumping bud：Explant obtained by step (1) is inoculated in culture medium M1, first light culture 8-10 days, then illumination Culture 80-90 days, until Multiple Buds are produced, and the bud height of Multiple Buds is not less than 2cm；The composition of the culture medium M1 is：MS bases Basal culture medium, supplement 1.0-2.0mg/L TDZ, 0.1-0.2mg/L IAA；(3) Multiple Buds switching culture：Multiple Buds obtained by step (2) are forwarded in culture medium M2, cultivated 15-25 days, until clump Sprout and grow up to the high Strawberry seedlings of 3-5cm, then the Strawberry seedlings are forwarded in culture medium M3, carry out culture of rootage, produce Strawberry detoxic seedling；The culture medium M2 is MS minimal mediums, and the culture medium M3 is 1/2MS minimal mediums.
- A kind of 2. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 1, it is characterised in that：Step (1) in, the sterilising conditions of the bud are：First with 75% alcohol sterilize 18-25s, then with 0.1% mercuric chloride solution sterilizing 12- 15min。
- A kind of 3. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 2, it is characterised in that：Step (1) in, the sterilising conditions of the bud are：First with 75% alcohol sterilize 20s, then with 0.1% mercuric chloride solution sterilize 13min.
- A kind of 4. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 1, it is characterised in that：Step (2) in, the explant is inoculated in culture medium M1, first light culture 9 days, then illumination cultivation 80 days.
- A kind of 5. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 1, it is characterised in that：Step (2) in, the condition of culture is 2000-3000LX of intensity of illumination, light irradiation time 16h/ days, 24-26 DEG C of cultivation temperature.
- A kind of 6. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 1, it is characterised in that：Step (3) in, the Multiple Buds are forwarded in culture medium M2, are cultivated 20 days.
- A kind of 7. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 1, it is characterised in that：Step (3) in, the condition of culture that the Multiple Buds are in culture medium M2 or the Strawberry seedlings are in culture medium M3 is：Intensity of illumination 2000-3000LX, light irradiation time 16h/ days, 24-26 DEG C of cultivation temperature.
- A kind of 8. method that strawberry detoxic seedling is cultivated using tissue culture technology according to claim 1, it is characterised in that：Step (1) in, the kind of the strawberry is selected from a liquor-saturated chivalrous, snow younger sister, chapter Ji, more beautiful for any one.
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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 |
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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 |
Non-Patent Citations (2)
| Title |
|---|
| "草莓"拉松7号"花药培养研究";任建宏;《安徽农业科学》;20101231;第38卷(第5期);第2250-2253页，1.2 方法，1.2.2 草莓花蕾的预处理和消毒方法，1.2.4 培养条件，摘要，2.4.4 对组培苗生根的影响 * |
| 植物生长调节剂TDZ在草莓花药培养中的应用;王敬东;《安徽农业科学》;20111231;第39卷(第5期);第2588-2590 * |
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