CN105850731B - A kind of culture medium and method of thickleaf spruce mature zygotic embryos callus induction - Google Patents
A kind of culture medium and method of thickleaf spruce mature zygotic embryos callus induction Download PDFInfo
- Publication number
- CN105850731B CN105850731B CN201610212622.XA CN201610212622A CN105850731B CN 105850731 B CN105850731 B CN 105850731B CN 201610212622 A CN201610212622 A CN 201610212622A CN 105850731 B CN105850731 B CN 105850731B
- Authority
- CN
- China
- Prior art keywords
- culture medium
- seed
- callus
- zygotic embryos
- thickleaf spruce
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
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
- A01H4/001—Culture apparatus for tissue culture
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to a kind of culture medium of thickleaf spruce mature zygotic embryos callus induction, it is characterised in that the callus inducing medium is the D 1mgL of S culture mediums+2,4‑1+6‑BA2mg·L‑1+KT 4mg·L‑1, the culture medium contains 34gL‑1Sucrose, 8gL‑1Agar, 0.5gL‑1Caseinhydrolysate, pH 5.8.The invention further relates to a kind of abductive approach of thickleaf spruce mature zygotic embryos callus.Advantages of the present invention is embodied in：For inductivity up to more than 80%, the method can be to establish stable thickleaf spruce somatic embryo occur system to lay the foundation, and have certain reference role to related scientific research, technical staff.
Description
Technical field
The present invention relates to Pinaceae spruce somatic embryo occur research field, and in particular to a kind of thickleaf spruce into
The culture medium and abductive approach of ripe zygotic embryo callus.
Background technology
Loose China fir class confierophyte is the important species of production of forestry and afforestation practice, but uses conventional vegetative propagation technique
It is difficult to take root, causes the utilization of its superior genotypes to be restricted.Then, people attempt real by the Development of Somatic Embryogenesis
The fast breeding of existing coniferous tree.1985, Hakman etc. is reported first to be closed using the prematurity of European spruce (Picea abies)
Sub- embryo success inducing somatic embryogenesis simultaneously obtains regeneration plant.Hereafter, the inducing of somatic embryos of coniferous trees, ripe and plant be again
The research of raw process has made great progress, and it is coniferous tree fine breed gene type scale to generally believe somatic embryo in the world
Change and expand numerous optimal path.
Thickleaf spruce (P.crassifolia) is China's eastern Tibetan north edge endemic tree, belongs to the evergreen pin of Picea
Leaf arbor.Be distributed mainly on Qilian Mountain Area, Qinghai (to the east of Dulan, west incline to the north of mountain), Gansu (Hexi Corridor and Jingyuan, Yuzhong,
Xiahe, Jone, Zhouqu County), Ningxia (Helan Mountain, Liu Pan Shan), the area of Daqingshan Mountains height above sea level 1600-3800 rice.Its material
It is excellent, drought resistance is stronger, be East of Qinghai Province, Northern Kansu mountain area and the excellent reproducting tree species of Qilian Mountain Area, and this area is gloomy
The climax of woods succession and the constructive species of zonal vegetation.
The content of the invention
The purpose of the present invention is to be directed to deficiency of the prior art, there is provided one kind is induced using thickleaf spruce mature zygotic embryos
The culture medium and method of callus.
To achieve the above object, the invention discloses following technical scheme：
A kind of culture medium of thickleaf spruce mature zygotic embryos callus induction, the callus inducing medium are trained for S
Support base+2,4-D 1mgL-1+6-BA 2mg·L-1+KT 4mg·L-1, the culture medium contains 34gL-1Sucrose, 8gL-1Fine jade
Fat, 0.5gL-1Caseinhydrolysate, pH 5.8.
Another object of the present invention is to disclose a kind of abductive approach of thickleaf spruce mature zygotic embryos callus, including such as
Lower step：
(1) culture medium is prepared：Minimal medium is configured by S culture medium prescriptions, then adds 1mgL-12,4-D,
2mg·L-16-BA, 4mgL-1KT and 0.5gL-1Caseinhydrolysate, and 34gL-1Sucrose and 8gL-1Fine jade
Fat, pH 5.8 are standby after packing sterilizing.
(2) explant sterilizes：After thickleaf spruce seed is soaked into 10-15h in water, kind of a skin is peelled off with tweezers；Then, use
75% alcohol carries out surface sterilization to the seed of peeling；0.1%HgCl is used again2Seed depth is sterilized, then rushed with sterilized water
Wash；Finally, aseptically, the surface of the seed moisture is blotted with aseptic filter paper.
(3) embryo separates：Aseptically, the tip portion of seed embryo root segment is fallen with sterilized fine-pointed forceps sub-folder,
Then pressed with tweezers and obtain embryo in the middle part of seed；
(4) it is inoculated with and cultivates：Aseptically, by the parallel surface for being inoculated in culture medium of embryo, 25 DEG C of conditions are placed in
Lower light culture；
(5) propagation of callus：After Callus formation, it is transferred in inducing culture, carries out Multiplying culture.
As a kind of further improvement project, in the step (2), thickleaf spruce seed is soaked into 12h in water.
As a kind of further improvement project, in the step (2), surface sterilization is carried out to the seed of peeling with alcohol
Time be 5min.
As a kind of further improvement project, in the step (2), with aseptic water washing 5 times, each 5min.
The culture medium and abductive approach of a kind of thickleaf spruce mature zygotic embryos callus disclosed by the invention, have following
Beneficial effect：
For the inductivity of the present invention up to more than 80%, the method can be to establish stable thickleaf spruce somatic embryo occur system
Lay the foundation, there is certain reference role to related scientific research, technical staff.
Embodiment
With reference to embodiment, the invention will be further described.
Embodiment 1
A kind of culture medium of thickleaf spruce mature zygotic embryos callus induction, the callus inducing medium are trained for S
Support base+2,4-D 8mgL-1+6-BA 0.5mg·L-1+KT 2mg·L-1, the culture medium contains 34gL-1Sucrose, 8gL-1Fine jade
Fat, 0.5gL-1Caseinhydrolysate, pH 5.8.The culture medium callus induction rate average out to 44.4%.
Embodiment 2
A kind of culture medium of thickleaf spruce mature zygotic embryos callus induction, the callus inducing medium are trained for S
Support base+2,4-D 4mgL-1+6-BA 4mg·L-1+KT 0.5mg·L-1, the culture medium contains 34gL-1Sucrose, 8gL-1Fine jade
Fat, 0.5gL-1Caseinhydrolysate, pH 5.8.The culture medium callus induction rate average out to 51.5%.
Embodiment 3
A kind of culture medium of thickleaf spruce mature zygotic embryos callus induction, the callus inducing medium are trained for S
Support base+2,4-D 1mgL-1+6-BA 2mg·L-1+KT 4mg·L-1, the culture medium contains 34gL-1Sucrose, 8gL-1Fine jade
Fat, 0.5gL-1Caseinhydrolysate, pH 5.8.The culture medium callus induction rate average out to 80.1%.
A kind of abductive approach of thickleaf spruce mature zygotic embryos callus, comprises the following steps：
(1) culture medium is prepared：Minimal medium is configured by S culture medium prescriptions, then adds 1mgL-12,4-D,
2mg·L-16-BA, 4mgL-1KT and 0.5gL-1Caseinhydrolysate, and 34gL-1Sucrose and 8gL-1Fine jade
Fat, pH 5.8 are standby after packing sterilizing.
(2) explant sterilizes：Thickleaf spruce seed is soaked in water (preferably 12h) after 10-15h, kind is peelled off with tweezers
Skin；Then, surface sterilization 5min is carried out to the seed of peeling with 75% alcohol；0.1%HgCl is used again2Seed depth is sterilized,
Then aseptic water washing is used 5 times, each 5min；Finally, aseptically, the surface of the seed moisture is blotted with aseptic filter paper.
(3) embryo separates：Aseptically, the tip portion of seed embryo root segment is fallen with sterilized fine-pointed forceps sub-folder,
Then pressed with tweezers and obtain embryo in the middle part of seed；
(4) it is inoculated with and cultivates：Aseptically, by the parallel surface for being inoculated in culture medium of embryo, 25 DEG C of conditions are placed in
Lower light culture；
(5) propagation of callus：After Callus formation, it is transferred in inducing culture, carries out Multiplying culture.
Described above is only the preferred embodiment of the present invention, rather than its limitations；It should be pointed out that although with reference to above-mentioned each
The present invention is described in detail embodiment, it will be understood by those within the art that, it still can be to above-mentioned each
Technical scheme described in embodiment is modified, or carries out equivalent substitution to which part or all technical characteristic；And this
A little modifications and replacement, do not make the essence of corresponding technical scheme depart from the scope of various embodiments of the present invention technical scheme.
Claims (5)
1. a kind of culture medium of thickleaf spruce mature zygotic embryos callus induction, it is characterised in that the callus induction is trained
It is S culture medium+2,4-D 1mgL to support base-1+6-BA 2mg·L-1+KT 4mg·L-1, the culture medium contains 34gL-1Sucrose,
8g·L-1Agar, 0.5gL-1Caseinhydrolysate, pH 5.8.
2. a kind of abductive approach of thickleaf spruce mature zygotic embryos callus, it is characterised in that comprise the following steps：
(1) culture medium is prepared：Minimal medium is configured by S culture medium prescriptions, then adds 1mgL-12,4-D, 2mgL-1
6-BA, 4mgL-1KT and 0.5gL-1Caseinhydrolysate, and 34gL-1Sucrose and 8gL-1Agar, pH are
5.8, it is standby after packing sterilizing；
(2) explant sterilizes：After thickleaf spruce seed is soaked into 10-15h in water, kind of a skin is peelled off with tweezers；Then, with 75%
Alcohol carries out surface sterilization to the seed of peeling；0.1%HgCl is used again2Seed depth is sterilized, then uses aseptic water washing；Most
Afterwards, aseptically, the surface of the seed moisture is blotted with aseptic filter paper；
(3) embryo separates：Aseptically, the tip portion of seed embryo root segment is fallen with sterilized fine-pointed forceps sub-folder, then
Embryo is obtained with tweezers pressure seed middle part；
(4) it is inoculated with and cultivates：Aseptically, it is dark under the conditions of being placed in 25 DEG C by the parallel surface for being inoculated in culture medium of embryo
Culture；
(5) propagation of callus：After Callus formation, it is transferred in inducing culture, carries out Multiplying culture.
A kind of 3. abductive approach of thickleaf spruce mature zygotic embryos callus according to claim 2, it is characterised in that
In the step (2), thickleaf spruce seed is soaked into 12h in water.
A kind of 4. abductive approach of thickleaf spruce mature zygotic embryos callus according to claim 2, it is characterised in that
In the step (2), the time for carrying out surface sterilization to the seed of peeling with alcohol is 5min.
A kind of 5. abductive approach of thickleaf spruce mature zygotic embryos callus according to claim 2, it is characterised in that
In the step (2), with aseptic water washing 5 times, each 5min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610212622.XA CN105850731B (en) | 2016-04-07 | 2016-04-07 | A kind of culture medium and method of thickleaf spruce mature zygotic embryos callus induction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610212622.XA CN105850731B (en) | 2016-04-07 | 2016-04-07 | A kind of culture medium and method of thickleaf spruce mature zygotic embryos callus induction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105850731A CN105850731A (en) | 2016-08-17 |
| CN105850731B true CN105850731B (en) | 2018-01-09 |
Family
ID=56636073
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610212622.XA Active CN105850731B (en) | 2016-04-07 | 2016-04-07 | A kind of culture medium and method of thickleaf spruce mature zygotic embryos callus induction |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105850731B (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104304034B (en) * | 2014-11-11 | 2017-01-11 | 新疆林科院造林治沙研究所 | Induction culture method and special induction culture medium for somatic embryo calluses of picea schrenkiana |
-
2016
- 2016-04-07 CN CN201610212622.XA patent/CN105850731B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105850731A (en) | 2016-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107047320B (en) | A kind of bigflower centranthera root method for tissue culture | |
| CN103125244A (en) | Cutting propagation method of golden camellia plants | |
| CN106106163A (en) | A kind of iris cultivates propagation method | |
| CN105010143B (en) | A kind of extracorporeal culturing method of Chinese catalpa | |
| CN101595824B (en) | Rapid in-vitro seedling raising method by utilizing sandalwood seed embryo | |
| CN106922534A (en) | A kind of method that tissue cultures quickly breed China fir | |
| CN104920228A (en) | Lilium duchartrei franch tissue culture rapid propagation and separation preservation method | |
| CN104041412A (en) | Rapid propagation method for tissue culture of Guizhou hemiboea cavaleriei | |
| CN105766654B (en) | A kind of Nanchuan jackfruit method for tissue culture | |
| CN109220790A (en) | A kind of in vitro outer breeding method of red fruit ginseng | |
| CN103404437B (en) | Novel method for tissue culture rapid propagation of acer paimatum | |
| CN103141388A (en) | Tissue culture method for ornithogalum caudatum | |
| CN105104206B (en) | See the blue or green in-vitro conservation method of blood | |
| CN104823846B (en) | The method for quickly breeding of Zhejiang Anoectochilus nefiliforme (Nakai) hara seedling | |
| CN105850741A (en) | Rapid propagation and in-vitro preservation method of coniogramme japonica (Thunberg) diels | |
| CN104488722B (en) | A kind of quick breeding method for tissue culture of South America crutch flower | |
| CN106613079A (en) | Method for producing pinellia-ternate seed stems | |
| CN103392593A (en) | Method for rapid tissue culture propagation of Dendrobium huoshanense seedlings | |
| CN105918119B (en) | A kind of method of the in vitro highly efficient regeneration of chu chrysanthemum blade | |
| CN105309312B (en) | A kind of method of Chinese purpleflower stonecrop herb tissue-culturing quick-propagation | |
| CN103141384A (en) | Rapid tissue culture propagation method of pot chrysanthemum cultivars | |
| CN105850731B (en) | A kind of culture medium and method of thickleaf spruce mature zygotic embryos callus induction | |
| CN104920218A (en) | Pararuellia delavayana reproduction method | |
| CN105165609A (en) | Method for efficiently acquiring gerbera jamesonii detoxification seedling | |
| CN104488710B (en) | A kind of method of Calotropis gigantea tissue cultures |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 733000 No. 1856, Qilian Avenue, Liangzhou District, Gansu, Wuwei Applicant after: Gansu Prov. Sand Control Research Inst. Address before: 730070 Gansu city of Lanzhou province Anning District North Riverside Road No. 390 Applicant before: Gansu Prov. Sand Control Research Inst. |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |