CN108012924A - A kind of Black Box Tracing tender shoots method for inducing and cultivating - Google Patents
A kind of Black Box Tracing tender shoots method for inducing and cultivating Download PDFInfo
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- CN108012924A CN108012924A CN201711414748.6A CN201711414748A CN108012924A CN 108012924 A CN108012924 A CN 108012924A CN 201711414748 A CN201711414748 A CN 201711414748A CN 108012924 A CN108012924 A CN 108012924A
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- black box
- tender shoots
- inducing
- box tracing
- 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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- Developmental Biology & Embryology (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of Black Box Tracing tender shoots method for inducing and cultivating, include the following steps：The selected Black Box Tracing plant to be cultivated, the spray of growth period clip stalwartness no disease and pests harm, through aseptic process, it is inoculated on inducing culture, Fiber differentiation is carried out in the case where being adapted to light and temperature condition, by callus induction, inducing clumping bud, forms sterile in vitro bud, Initial culture system under aseptic condition is established, basic material is provided for the squamous subculture of next step.Material of the present invention is simple and easy to get, and each link is easy to operate, and condition of culture is controllable, and reproducibility of results is strong, and cost is low, beneficial to promoting and putting into production, to promoting Black Box Tracing good seed is numerous soon to have important practice significance.
Description
Technical field
The present invention relates to agricultural and flowers and trees cultivation field, and in particular to a kind of Black Box Tracing (Aronia
MeLanocarpa) sterile induction Multiple Buds method.
Background technology
Black Box Tracing is a seeds for rose family gland Aronia, is the seeds introduced from foreign countries, referred to as black fruit
Sorbus alnifloria, black fruit, the not old certain kind of berries, wild cherry certain kind of berries etc., machaka, sets high 1.5-2.5m, and plexi is tree-like, is planted for afforestation, berry
Training.Black Box Tracing is adapted to the weather of the Northeast's winter cold, and cultivated area constantly expanded in recent years, and seedling lacks, especially
The pure seedling of its kind is rare.It is to solve seedling demand greatly, in short term that seedling is quickly produced using plant tissue culture technique
It is irreproducible to go out to need the optimal selection of problem enough.Since Black Box Tracing is introduced, the domestic time is shorter, and tissue culture is real
It is particularly important that being in blank situation with technology, especially established this link of sterile induction system, primary Fiber differentiation are that subculture is fast
Numerous basis is, it is necessary to which a large number of experiments can just filter out best-of-breed technology measure.
The peasant of black fruit Sorbus alnifloria plants in the Northeast at present and enterprise is more, it is necessary to which good seed amount is estimated up to several ten million plants
More than, market needs cannot be met by providing seedling by plant division and cottage method, and seedling shortage constrains black fruit Sorbus alnifloria industry development.
The content of the invention
For many years, in experimental basis, the present invention provides a kind of Black Box Tracing tender shoots method for inducing and cultivating, it is ensured that
The development of Black Box Tracing tissue culture sprout quick propagating technology, provides sufficient sterilizable material, when saving substantial amounts of grope for squamous subculture
Between.
To achieve the above object, the technical solution taken of the present invention is：
A kind of Black Box Tracing tender shoots method for inducing and cultivating, includes the following steps：
Determine to expand numerous Black Box Tracing plant region, from wherein selecting, root system is strong, limb is sturdy, result quantities
Greatly, the plant of no disease and pests harm elects collection explant elite stand as, in the month in growth period 4-8 or rest period, takes a healthy and strong branch long shoot section,
After completing pretreatment, in superclean bench, under aseptic condition, tender shoots is taken, few band xylem, is placed in alcohol and sterilizes 20 seconds
Afterwards, immersion in glutaraldehyde solution is placed in, with aseptic water washing, is inoculated on culture medium, induction training is carried out in Intelligent culture case
Support 20 days.
Preferably, the time gathers the healthy and strong branch long shoot section in the morning, uses polybag moisturizing bag after shear offing immediately
Install, quickly take laboratory and carry out aseptic process.
Preferably, specifically comprise the following steps：
S1, determine to expand numerous Black Box Tracing plant region, and from wherein selecting, root system is strong, limb is sturdy, result
Amount is big, the plant of no disease and pests harm elects collection explant elite stand as, in 5-8 months, in the plant strain growth phase, selects semi-lignified
Healthy and strong branch, clip when moisture is full in the morning, when flushing branch 1 is small under flowing water, puts in suds and soaks 10 minutes, then put
Rinsed under flowing water after twenty minutes, with aseptic water washing branch, be cut into 3 centimeter length short sections, put in sterile water and soak, move into ultra-clean work
Make pre-sterilization 30 minutes under platform ultraviolet light；
S2, in superclean bench, under aseptic condition, clip tender shoots, few band branch xylem, being placed in volume fraction is
After being sterilized 20 seconds in 70% alcohol, it is put into 20g/L glutaraldehyde solutions and soaks 10 minutes；With aseptic water washing 3 times, wood is cut
Matter part, peels off outer blade, and tender shoots is inoculated into inducing culture, is inserted into 0.3-0.5 centimetres of depth of culture medium, upright to insert
Enter, every bottle of tender shoots；
Blake bottle after inoculation, be placed in Intelligent culture case and carry out Fiber differentiation 20 days by S3, it is seen that tender shoots incision is given birth to
Go out callus, tender shoots base portion grows 3-5 sprouting, hanks work(and induces the blake bottle of Multiple Buds or stem section for rolling bottle training
Support, into the squamous subculture stage；Condition of culture is arranged to day temperature control at 22-24 DEG C, and night temperatures control is at 20 DEG C, light
According to strength control in 1500Lx, when daily illumination 12 is small, daytime is to light, night lucifuge.
Preferably, the inducing culture is prepared by the following method：
Using MS minimal mediums, sucrose 30g/L, agar powder 6g/L, 6-BA (6 benzyl purine) 2.0g/L, NAA are added
(methyl α-naphthyl acetate) 0.2g/L, pH6.5, is filled into 50mL blake bottles, and culture medium thickness is controlled in 1cm.
Preferably, to be gathered the plant of explant and need to shift to an earlier date 15d and not spray medicine and fertilizer, normal watering management.
The invention has the advantages that：
Material of the present invention is simple and easy to get, and each link is easy to operate, and condition of culture is controllable, and reproducibility of results is strong, and cost is low,
Beneficial to promoting and put into production；Fiber differentiation is the key link of Plant Tissue Breeding, gropes that time-consuming, and sterile working needs ripe
Practice personnel to carry out, under the guidance of this inventive step, aseptic seedling system can be entered in 2 months and start link.The invention uses model
Big, technical specification is enclosed, beneficial to large-scale production, element task has been carried out for squamous subculture, culture of rootage.Personal and company
Use, beneficial to factorial praluction, can also carry on fast numerous experiment.It is in short supply for solving the Northeast's Black Box Tracing seedling
Problem, there is provided technical guarantee.
Embodiment
In order to which objects and advantages of the present invention are more clearly understood, the present invention is carried out with reference to embodiments further
Describe in detail.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to limit this hair
It is bright.
Embodiment 1
A kind of Black Box Tracing tender shoots method for inducing and cultivating, includes the following steps：
Numerous Black Box Tracing plant will be expanded and elect collection explant elite stand as, 20 centimeter length of top tip is sheared in growth period
Branch section, rinses under flowing water, is cut into 3 centimeter length sections, in superclean bench, under aseptic condition, and clip tender shoots, few band branch wood
Matter portion, puts in alcohol and sterilizes 20 seconds, is subsequently placed into glutaraldehyde solution and soaks 10 minutes, with aseptic water washing, is inoculated into culture
Base, MS+ sucrose 30g/L+ agar powders 6g/L+6-BA (6 benzyl purine) 2.0g/L+NAA (methyl α-naphthyl acetate) 0.2g/L, in Intelligent culture
Fiber differentiation is carried out in case 20 days.
Embodiment 2
A kind of Black Box Tracing tender shoots method for inducing and cultivating, includes the following steps：
Numerous Black Box Tracing plant will be expanded and elect collection explant elite stand as, 1 year raw branch 30 is sheared in rest period
Centimetre long shoot section, moves into water cutting culture in greenhouse, plastic foil button canopy heat and moisture preserving, shear offs after sprouting sprouts, in flowing water
Lower flushing, immerses in sterile water and is placed in superclean bench, under aseptic condition, strip tender shoots, and few band xylem, puts in alcohol and disappear
Poison 20 seconds, is subsequently placed into glutaraldehyde solution and soaks 10 minutes, with aseptic water washing, be inoculated into culture medium, MS+ sucrose 30g/L+
Agar powder 6g/L+6-BA (6 benzyl purine) 2.0g/L+NAA (methyl α-naphthyl acetate) 0.2g/L, carries out Fiber differentiation in Intelligent culture case
20 days.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the principle of the present invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (5)
1. a kind of Black Box Tracing tender shoots method for inducing and cultivating, it is characterised in that include the following steps：
Determine to expand numerous Black Box Tracing plant region, from wherein selecting, root system is strong, limb is sturdy, result quantities are big, nothing
The plant of pest and disease damage elects collection explant elite stand as, in the month in growth period 4-8 or rest period, takes a healthy and strong branch long shoot section, completes
After pretreatment, in superclean bench, under aseptic condition, tender shoots is taken, few band xylem, is placed in alcohol after sterilizing 20 seconds, puts
In being soaked in glutaraldehyde solution, with aseptic water washing, it is inoculated on culture medium, Fiber differentiation 20 is carried out in Intelligent culture case
My god.
A kind of 2. Black Box Tracing tender shoots method for inducing and cultivating as claimed in claim 1, it is characterised in that the stalwartness branch
The time gathers bar long shoot section in the morning, is packaged immediately with polybag moisturizing after shear offing, and quickly takes laboratory and carries out nothing
Bacterium is handled.
A kind of 3. Black Box Tracing tender shoots method for inducing and cultivating as claimed in claim 1, it is characterised in that specifically include as
Lower step：
S1, determine to expand numerous Black Box Tracing plant region, from wherein select root system it is strong, limb is sturdy, result quantities are big,
The plant of no disease and pests harm elects collection explant elite stand as, in 5-8 months, in the plant strain growth phase, selects the stalwartness of semi-lignified
Branch, clip when moisture is full in the morning, when flushing branch 1 is small under flowing water, puts in suds and soaks 10 minutes, then release water
Lower flushing after twenty minutes, with aseptic water washing branch, is cut into 3 centimeter length short sections, puts in sterile water and soak, and moves into superclean bench
Pre-sterilization 30 minutes under ultraviolet light；
S2, in superclean bench, under aseptic condition, clip tender shoots, few band branch xylem, it is 70% to be placed in volume fraction
After being sterilized 20 seconds in alcohol, it is put into the glutaraldehyde solution of 20g/L and soaks 10 minutes；With aseptic water washing 3 times, lignifying is cut
Part, peels off outer blade, and tender shoots is inoculated into inducing culture, is inserted into 0.3-0.5 centimetres of depth of culture medium, upright to be inserted into,
Every bottle of tender shoots；
Blake bottle after inoculation, be placed in Intelligent culture case and carry out Fiber differentiation 20 days by S3, it is seen that tender shoots incision is born more
Injured tissue, tender shoots base portion grow 3-5 sprouting, hank that work(induces Multiple Buds or the blake bottle of stem section is used for spinner culture, into
Enter the squamous subculture stage；Condition of culture is arranged to day temperature control at 22-24 DEG C, and night temperatures are controlled at 20 DEG C, and illumination is strong
Degree control is in 1500Lx, and when daily illumination 12 is small, daytime is to light, night lucifuge.
A kind of 4. Black Box Tracing tender shoots method for inducing and cultivating as claimed in claim 1, it is characterised in that the induction training
Foster base is prepared by the following method：
Using MS minimal mediums, sucrose 30g/L, agar powder 6g/L, 6-BA 2.0g/L, NAA 0.2g/L, pH6.5 are added, is filled
It is fitted into 50mL blake bottles, culture medium thickness is controlled in 1cm.
5. a kind of Black Box Tracing tender shoots method for inducing and cultivating as claimed in claim 1, it is characterised in that to be gathered outer
The plant of implant need to shift to an earlier date 15d and not spray medicine and fertilizer, normal watering management.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711414748.6A CN108012924A (en) | 2017-12-14 | 2017-12-14 | A kind of Black Box Tracing tender shoots method for inducing and cultivating |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711414748.6A CN108012924A (en) | 2017-12-14 | 2017-12-14 | A kind of Black Box Tracing tender shoots method for inducing and cultivating |
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| CN108012924A true CN108012924A (en) | 2018-05-11 |
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| CN201711414748.6A Pending CN108012924A (en) | 2017-12-14 | 2017-12-14 | A kind of Black Box Tracing tender shoots method for inducing and cultivating |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108901847A (en) * | 2018-07-24 | 2018-11-30 | 江苏农牧科技职业学院 | A kind of propagation method of Black Box Tracing and kit for this method |
| CN109463283A (en) * | 2018-12-26 | 2019-03-15 | 武威茂瑞特色树种技术推广有限公司 | Black Box Tracing tender shoots method for inducing and cultivating |
| CN109479541A (en) * | 2018-12-26 | 2019-03-19 | 武威凯弘利特色农产品开发有限公司 | A kind of Black Box Tracing cuttage breeding method |
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| KR20140037510A (en) * | 2012-09-19 | 2014-03-27 | 농업회사법인 호트팜 주식회사 | Multiple propagation methods of in vitro plantlets derived from node cultures of aronia using tissue culture techniques |
| CN106069790A (en) * | 2016-08-24 | 2016-11-09 | 济南浩隆生物科技有限公司 | A kind of Black Box Tracing open tissue cultivates fast seedling-cultivating method |
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| KR20140037510A (en) * | 2012-09-19 | 2014-03-27 | 농업회사법인 호트팜 주식회사 | Multiple propagation methods of in vitro plantlets derived from node cultures of aronia using tissue culture techniques |
| CN106069790A (en) * | 2016-08-24 | 2016-11-09 | 济南浩隆生物科技有限公司 | A kind of Black Box Tracing open tissue cultivates fast seedling-cultivating method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108901847A (en) * | 2018-07-24 | 2018-11-30 | 江苏农牧科技职业学院 | A kind of propagation method of Black Box Tracing and kit for this method |
| CN109463283A (en) * | 2018-12-26 | 2019-03-15 | 武威茂瑞特色树种技术推广有限公司 | Black Box Tracing tender shoots method for inducing and cultivating |
| CN109479541A (en) * | 2018-12-26 | 2019-03-19 | 武威凯弘利特色农产品开发有限公司 | A kind of Black Box Tracing cuttage breeding method |
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Application publication date: 20180511 |