CN105613297B - Job's tears tissue cultivation rapid breeding method - Google Patents
Job's tears tissue cultivation rapid breeding method Download PDFInfo
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- CN105613297B CN105613297B CN201610048708.3A CN201610048708A CN105613297B CN 105613297 B CN105613297 B CN 105613297B CN 201610048708 A CN201610048708 A CN 201610048708A CN 105613297 B CN105613297 B CN 105613297B
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- job
- tears
- callus
- culture
- seedling
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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
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of Job's tears tissue cultivation rapid breeding method, this method comprises the following steps：Choose Job's tears explant to carry out disinfection processing, the Job's tears explant disinfected is put into Job's tears callus differential medium and carries out culture differentiation；After turning out Job's tears callus, then by Job's tears callus subculture medium carry out strengthen Job's tears callus squamous subculture；Then Job's tears dedifferentiation culture is carried out by Job's tears redifferential medium；Being broken up again by Job's tears makes Job's tears be cultivated when reaching seedling using Job's tears seedling culture medium；Job's tears culture of rootage is carried out by Job's tears root media after Job's tears seedling.This Job's tears tissue cultivation rapid breeding method callus induction differentiation efficiency height of the present invention, induction seedling effect are good, it is possible to achieve the preservation of Job's tears germ plasm resource, establish genetic conversion system and tissue cultural seedlings of free production, be worthy to be popularized.
Description
Technical field
The present invention relates to Job's tears tissue culture rapid propagating technology, belong to technical field of plant cultivation.
Background technology
Job's tears is grass family Coix (coix.L.) crop, and its seed (being commonly called as Semen Coicis) is nutritious, can make edible and medicine
With full category shares 10 kinds, and China reports that the category has a kind of 1 mutation, i.e. Chuan Gu and Job's tears altogether at present.Currently, the development of Job's tears industry
The problem of being primarily present has single varieties, disease are serious, yield is low etc., seriously restricts its development；Problem one be present more than solving
As in cereal crops practices well be exactly by its economical character import excellent genes its character is effectively changed
It is good, but the success of genetic transformation needs the foundation of high-efficiency regeneration system, and Plant Tissue Breeding is a basis in genetic transformation
And important link, therefore, establishing highly efficient regeneration genetic conversion system need to complete by plant tissue culture technique means.
Plant tissue's culture technique refers to plant aseptically, be organized into, cell, embryo, a system such as protoplast
It is classified as explant and Fiber differentiation is carried out on the culture medium manually prepared, when the condition of culture given is suitable, induces callus group
Knit, resting bud etc. produces, and forms the process of complete plant.The technology originates from 1902, is by German botanist
Based on Haberlandt proposes the theory of " plant cell has totipotency ", the theoretical proposition is Plant Tissue Breeding skill
Art development provides wide prospect, and it is multi-field to be widely used in flowers, crops, fruit tree, Germ-plasma resources protection etc. thereafter.
Plant tissue culture technique is one of maximally effective approach in current plant fast propagation, has that the breeding cycle is short, breeding coefficient is high,
It is easy to mass produce, the process has the advantage of low production cost, is also applied by maturation in important cereal crops, and cultivate
Go out large quantities of new varieties.The application of tissue cultures and regeneration research on corn lures explant with being just to be able to high-frequency
Callus is exported, accelerates Somatic Embryogenesis, promotes corn gene transformation.Therefore this technology is applied successfully to the heart of a lotus seed
Preservation, Establishment of Agrobacterium-Mediated Transformation System and tissue cultural seedlings of free production of Job's tears germ plasm resource etc. should all have important meaning
Justice.
In recent years, Job's tears tissue culture technique mainly breaks up to callus induction and again, flower pesticide tissue culture supports aspect
Work, the technologies such as low, the induction seedling effect difference of differentiation efficiency are induced to need for callus present in Job's tears tissue culture technique
Further optimization.Therefore, it is in Job's tears research to optimize Job's tears tissue culture technique and establish ripe Job's tears genetic conversion system
An important ring, laid the foundation for the research of Job's tears genomics and gene function.
The content of the invention
The purpose of the present invention is to realize the preservation of Job's tears germ plasm resource, establishing genetic conversion system and tissue cultural seedlings of free
Produce and a kind of Job's tears tissue cultivation rapid breeding method is provided.
Technical scheme is as follows：
This Job's tears tissue cultivation rapid breeding method of the present invention comprises the following steps：Job's tears explant is chosen first to be disappeared
Poison processing, the Job's tears explant disinfected is put into Job's tears callus differential medium and carries out culture differentiation；Turn out the heart of a lotus seed
After Job's tears callus, then by Job's tears callus subculture medium carry out strengthen Job's tears callus squamous subculture；Then by Job's tears again
Differential medium carries out Job's tears dedifferentiation culture；Being broken up again by Job's tears makes Job's tears be trained when reaching seedling using Job's tears seedling
Foster base is cultivated；Job's tears culture of rootage is carried out by Job's tears root media after Job's tears seedling.
Wherein, the reagent used is disinfected as mercuric chloride, sodium hypochlorite, alcohol.During using coix seed as explant, sterilization
Alcohol-pickled 1 min that it is first 75% with concentration by explant that processing method, which is, aseptic water washing 2 ~ 3 times, it is 0.1% to add concentration
Mercuric chloride soak 15 min or add the sodium hypochlorite that concentration is 10% and soak 20 min, then with aseptic water washing 3 ~ 5 times.
Wherein, Job's tears callus differential medium is MS+2,4-D 3.0mg/L+6-BA1.0mg/L.Job's tears callus subculture is trained
It is MS+2,4D 2.0mg/L+6-BA0.5mg/L to support base.Job's tears redifferential medium is MS+6-BA 4.0mg/L+2,4D
0.4mg/L.Job's tears seedling culture medium is MS+ZT 2.0mg/L+2,4D 0.4mg/L+KT 1.0mg/L.Job's tears root media
For 1/2MS+IBA0.5mg/L.
This Job's tears tissue cultivation rapid breeding method callus induction differentiation efficiency height of the present invention, induction seedling effect are good, can
With realize the preservation of Job's tears germ plasm resource, establish genetic conversion system and tissue cultural seedlings of free production, be worthy to be popularized.
Embodiment
With reference to embodiment, the present invention is described in further detail.
Embodiment 1：Carry out according to the following steps：
1st, it is explant and to be disinfected to choose coix seed, disinfect be with 75% alcohol-pickled 1min, it is sterile
Water rinses 2 times, adds 0.1% mercuric chloride and soaks 15 min, with aseptic water washing 3 times.
2nd, it is put into Job's tears callus differential medium with the Job's tears explant disinfected and carries out culture differentiation, the Job's tears is cured
It is MS+2,4-D 3.0mg/L+6-BA1.0mg/L to hinder differential medium.
3rd, by turning out Job's tears callus, Job's tears callus squamous subculture, the Job's tears callus subculture training of use are strengthened at the same time
It is MS+2,4D 2.0mg/L+6-BA 0.5mg/L to support base.
4th, by Job's tears callus subculture, carry out Job's tears and break up optimal culture again, the Job's tears redifferential medium used is MS+
6-BA 4.0mg/L+2,4D 0.4mg/L。
5th, broken up again by Job's tears, when Job's tears just can reach seedling, cultivated using Job's tears seedling culture medium, the heart of a lotus seed
Job's tears seedling culture medium MS+ZT 2.0mg/L+2,4D 0.4mg/L+KT 1.0mg/L.
6th, after by Job's tears seedling, it is possible to carry out Job's tears culture of rootage, Job's tears root media is 1/2MS+
IBA0.5mg/L。
Embodiment 2：Carry out according to the following steps：
1st, it is explant and to be disinfected to choose coix seed, disinfect be with 75% alcohol-pickled 1min, it is sterile
Water rinses 3 times, 10% sodium hypochlorite immersion 20min is added, with aseptic water washing 5 times.
2nd, it is put into Job's tears callus differential medium with the Job's tears explant disinfected and carries out culture differentiation, the Job's tears is cured
It is MS+2,4-D 3.0mg/L+6-BA1.0mg/L to hinder differential medium.
3rd, by turning out Job's tears callus, Job's tears callus squamous subculture, the Job's tears callus subculture training of use are strengthened at the same time
It is MS+2,4D 2.0mg/L+6-BA 0.5mg/L to support base.
4th, by Job's tears callus subculture, Job's tears dedifferentiation culture is carried out, the Job's tears redifferential medium used is MS+6-BA
4.0mg/L+2,4D 0.4mg/L。
5th, broken up again by Job's tears, when Job's tears just can reach seedling, cultivated using Job's tears seedling culture medium, the heart of a lotus seed
Job's tears seedling culture medium MS+ZT 2.0mg/L+2,4D 0.4mg/L+KT 1.0mg/L.
6th, after by Job's tears seedling, it is possible to carry out Job's tears culture of rootage, Job's tears root media is 1/2MS+
IBA0.5mg/L。
Certainly, the above is the concrete application example of the present invention, and the present invention also has other embodiments, all using equivalent
The technical scheme that replacement or equivalent transformation are formed, all falls within protection domain of the presently claimed invention.
Claims (6)
1. a kind of Job's tears tissue cultivation rapid breeding method, it is characterised in that comprise the following steps：Job's tears explant is chosen to carry out disinfection
Processing, the Job's tears explant disinfected is put into Job's tears callus differential medium and carries out culture differentiation；Turn out Job's tears
After callus, then by Job's tears callus subculture medium carry out strengthen Job's tears callus squamous subculture；Then divided again by Job's tears
Change culture medium and carry out Job's tears dedifferentiation culture；Being broken up again by Job's tears makes Job's tears use Job's tears seedling culture when reaching seedling
Base is cultivated；Job's tears culture of rootage is carried out by Job's tears root media after Job's tears seedling；The examination for disinfecting use
Agent is sodium hypochlorite；The Job's tears callus subculture medium is MS+2,4D 2.0mg/L+6-BA0.5mg/L.
2. Job's tears tissue cultivation rapid breeding method according to claim 1, it is characterised in that：Using coix seed as explant
When, alcohol-pickled 1 min that it is first 75% with concentration by explant that disinfection treatment method, which is, aseptic water washing 2 ~ 3 times, add dense
The sodium hypochlorite spent for 10% soaks 20 min, then with aseptic water washing 3 ~ 5 times.
3. Job's tears tissue cultivation rapid breeding method according to claim 1, it is characterised in that：The Job's tears callus differentiation culture
Base is MS+2,4-D 3.0mg/L+6-BA1.0mg/L.
4. Job's tears tissue cultivation rapid breeding method according to claim 1, it is characterised in that：The Job's tears redifferential medium
For MS+6-BA 4.0mg/L+2,4D 0.4mg/L.
5. Job's tears tissue cultivation rapid breeding method according to claim 1, it is characterised in that：The Job's tears seedling culture medium is
MS+ZT 2.0mg/L+2,4D 0.4mg/L+KT 1.0mg/L.
6. Job's tears tissue cultivation rapid breeding method according to claim 1, it is characterised in that：The Job's tears root media is
1/2MS+IBA0.5mg/L。
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610048708.3A CN105613297B (en) | 2016-01-26 | 2016-01-26 | Job's tears tissue cultivation rapid breeding method |
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| CN201610048708.3A CN105613297B (en) | 2016-01-26 | 2016-01-26 | Job's tears tissue cultivation rapid breeding method |
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| CN105613297A CN105613297A (en) | 2016-06-01 |
| CN105613297B true CN105613297B (en) | 2017-12-26 |
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| CN109169280B (en) * | 2018-09-20 | 2022-02-08 | 四川农业大学 | Preparation method of coix seed regenerated plant and culture medium used by preparation method |
| CN110024693B (en) * | 2019-05-07 | 2022-02-08 | 贵州大学 | Tissue culture and rapid propagation method by utilizing stem tip meristem of coix lacryma-jobi |
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| CN104335899B (en) * | 2014-10-08 | 2016-09-14 | 兰州大学 | Elymus nutans children's fringe isolated culture regeneration plant method |
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