CN107950392A - A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time - Google Patents
A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time Download PDFInfo
- Publication number
- CN107950392A CN107950392A CN201711179549.1A CN201711179549A CN107950392A CN 107950392 A CN107950392 A CN 107950392A CN 201711179549 A CN201711179549 A CN 201711179549A CN 107950392 A CN107950392 A CN 107950392A
- Authority
- CN
- China
- Prior art keywords
- subculture
- interval time
- regeneration plant
- culture
- culture medium
- 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.)
- Pending
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 provides a kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time,The subculture medium that this method uses is to contain 20~30g/L of soft white sugar,6 0.15~0.25mg/L of BA,8~10g/L of agar powder,0.35~0.45mg/L of NAA,Kanamycins (Kan) 0.1~0.2mg/L,PH value is 5.5~6.5 MS culture mediums,Culture medium dispensed loading amount is to load 70~80mL per 100mL triangular flasks,Regeneration plant is accessed in culture medium,At 20 ± 2 DEG C,2000~3000lx of intensity of illumination,After 7~10d being cultivated under conditions of photoperiod 12h/12h,Move into 2~8 DEG C of refrigerating boxes and place,Taken out when culture medium content drops to 10~20mL,Carry out subculture operation next time.In this approach subculture interval time can be made to reach 70~90d, be 2~3.5 times of cellar culture subculture interval time, the workload of subculture switching can be greatly reduced.
Description
Technical field
The present invention relates to belonging to biological technical field, more particularly to it is a kind of extend Chinese cabbage microspore embryoid regeneration plant after
For the method for interval time.
Technical background
Chinese cabbage is one of edible vegetable main in our people's life, deep for a long time to be liked by broad masses.For training
New germ plasm resource is educated, is bred as high-quality breeds of Chinese cabbage, conventional method needs, by hybridization, backcrossing and selfing separation, to select reason
The material thought usually needs several from generation to generation, and utilizes microspore culture, not only can quickly obtain dihaploid pure lines, also
The breeding time limit is substantially shorter, with utilizing isolated microspore culture technique phase in the research of Brassica Crops breeding both at home and abroad
After making a breakthrough, also application provides the foundation technology in practice for this.
It can shorten the breeding time limit using microspore culture, but exist in concrete operations and somatic embryogenesis is planted
Strain carries out the switching of multiple subculture and is further cultured for burying the asking of planting time so as to meet that reality escapes torridity summer to keep its activity
Topic, subculture switching will expend substantial amounts of manpower and time, so it is just very heavy to establish a kind of method for extending subculture interval time
Will, and extend subculture interval time then mainly the speed of growth and culture medium time of drying up carry out reality in the medium by delaying plant
It is existing.Found by studying, the factor such as component content, cultivation temperature, intensity of illumination and photoperiod of culture medium can give birth to plant
Long speed and culture medium withered time have an impact, and the present invention realizes by being optimized to factors above and extends subculture
The purpose of interval time.
The content of the invention
A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time, used subculture medium
For：Containing 20~30g/L of soft white sugar, 6-BA 0.15~0.25mg/L, NAA 0.35~0.45mg/L, 8~10g/L of agar powder,
Kanamycins (Kan) 0.1~0.2mg/L, pH value are 5.5~6.5 MS culture mediums；Culture medium dispensed loading amount is per 100mL triangles
It is bottled enter 70~80mL.
Further, concrete operation step is：(1) regeneration plant formed by embryoid is transferred into containing above-mentioned culture
In the triangular flask of base.
(2) 7~10d is cultivated under conditions of 20 ± 2 DEG C, 2000~3000lx of intensity of illumination, photoperiod 12h/12h.
(3) (2) step is inoculated with triangular flask 2~8 DEG C of refrigerating boxes of immigration after regeneration plant and placed, treat culture medium content
Taken out when dropping to 10~20mL, then carry out subculture switch over operation next time.
Further, subculture interval time can be made to reach 70~90d, is the 2~3.5 of cellar culture subculture interval time
Times.
Brief description of the drawings
Fig. 1 is the Chinese cabbage somatic embryogenesis plant subculture interval time table of comparisons.
Fig. 2 is the Chinese cabbage somatic embryogenesis plant subculture interval time table of comparisons.
Embodiment
Chinese cabbage microspore different genotype somatic embryogenesis plant is chosen, in the different subculture medium of four component contents
Middle culture, culture medium are respectively：
No. 1：Contain 20~30g/L of soft white sugar, 6-BA 0.35~0.45mg/L of 0.15~0.25mg/L, NAA, agar powder
8~10g/L, kanamycins (Kan) 0.1mg/L, pH value are 5.5~6.5 MS culture mediums；
No. 2：Contain 30~40g/L of soft white sugar, 6-BA 0.35~0.45mg/L of 0.25~0.35mg/L, NAA, agar powder
8~10g/L, kanamycins (Kan) 0.1mg/L, pH value are 5.5~6.5 MS culture mediums；
No. 3：Contain 20~30g/L of soft white sugar, 6-BA 0.25~0.35mg/L of 0.15~0.25mg/L, NAA, agar powder
6.5~7.5g/L, kanamycins (Kan) 0.2mg/LpH value are 5.5~6.5 MS culture mediums；
No. 4：Contain 30~40g/L of soft white sugar, 6-BA 0.25~0.35mg/L of 0.25~0.35mg/L, NAA, agar powder
6.5~7.5g/L, kanamycins (Kan) 0.1mg/L, pH value are 5.5~6.5 MS culture mediums；
Culture medium dispensed loading amount is to load 70~80mL per 100mL triangular flasks, after regeneration plant is transferred to culture medium, 20 ±
After cultivating 7d~10d under conditions of 2 DEG C, 2000~3000lx of intensity of illumination, photoperiod 12h/12h, 2~8 DEG C of refrigerating boxes are moved into
Middle placement, (such as occur culture medium is dry and cracked will seriously to be taken out in advance) is taken out when culture medium content drops to 10~20mL.Statistics
Culture medium is transferred to the time taken out from refrigerator from regeneration plant, the results are shown in Table 1,
Choose Chinese cabbage microspore different genotype somatic embryogenesis plant, select subculture medium be containing soft white sugar 20~
30g/L, 6-BA0.15~0.25mg/L, NAA0.35~0.45mg/L, 8~10g/L of agar powder, kanamycins (Kan) 0.1mg/
L, pH value are 5.5~6.5 MS culture mediums, are cultivated respectively under three different condition of culture, condition is respectively：
A. 20 ± 2 DEG C of temperature, 2000~3000lx of intensity of illumination, photoperiod 12h/12h, culture 7d~10d move into 2~8
Place in DEG C refrigerating box, taken out when culture medium content drops to 10~20mL.
B. 25 ± 3 DEG C of temperature, 3000~4000lx of intensity of illumination, photoperiod 16h/8h, culture 7d~10d move into 2~8 DEG C
Place in refrigerating box, taken out when culture medium content drops to 10~20mL.
C. 25 ± 3 DEG C of temperature, 3000~4000lx of intensity of illumination, photoperiod 16h/8h, room is placed on after cultivating 7d~10d
Under the conditions of temperature, stop culture when culture medium content drops to 10~20mL.
Count from regeneration plant and transfer culture medium to the time that culture is taken out or stopped under room temperature from refrigerator, join
Examine that the visible condition of culture A of attached drawing 2 is best to the prolongation effect of subculture interval time, and interval time can reach 70~90d, be culture
2~3.5 times of condition C subculture interval time.
Using technical solutions according to the invention, or those skilled in the art is under the inspiration of technical solution of the present invention,
Similar technical solution is designed, and reaches above-mentioned technique effect, is to fall into protection scope of the present invention.
Claims (3)
- A kind of 1. method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time, it is characterised in that：It is used Subculture medium is：Containing 20~30g/L of soft white sugar, 6-BA0.15~0.25mg/L, agar powder 8~10g/L, NAA0.35~ 0.45mg/L, kanamycins (Kan) 0.1~0.2mg/L, pH value are 5.5~6.5 MS culture mediums；Culture medium dispensed loading amount is every 100mL triangular flasks load 70mL~80mL.
- 2. a kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time according to claim 1, It is characterized in that：Concrete operation step is as follows,(1) regeneration plant formed as embryoid is transferred in the triangular flask containing culture medium described in claim 1；(2) 7~10d is cultivated under conditions of 20 ± 2 DEG C, 2000~3000lx of intensity of illumination, photoperiod 12h/12h；(3) (2) step is inoculated with triangular flask 2~8 DEG C of refrigerating boxes of immigration after regeneration plant and placed, treat that culture medium content declines Taken out during to 10~20mL, then carry out subculture switch over operation next time.
- 3. a kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time according to claim 1, It is characterized in that：Subculture interval time is reached 70~90d, be 2~3.5 times of cellar culture subculture interval time.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711179549.1A CN107950392A (en) | 2017-11-09 | 2017-11-09 | A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711179549.1A CN107950392A (en) | 2017-11-09 | 2017-11-09 | A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107950392A true CN107950392A (en) | 2018-04-24 |
Family
ID=61961747
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711179549.1A Pending CN107950392A (en) | 2017-11-09 | 2017-11-09 | A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107950392A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283589A (en) * | 2012-02-24 | 2013-09-11 | 河北农业大学 | Method for prolonging subculturing interval time of tissue culture seedlings of apple |
-
2017
- 2017-11-09 CN CN201711179549.1A patent/CN107950392A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283589A (en) * | 2012-02-24 | 2013-09-11 | 河北农业大学 | Method for prolonging subculturing interval time of tissue culture seedlings of apple |
Non-Patent Citations (4)
| Title |
|---|
| 严慧玲等: "甘蓝显性雄性不育材料离体保存技术的研究", 《热带农业科学》 * |
| 朱彦涛: "油菜小孢子培养和DH系繁殖研究", 《中国优秀硕博士学位论文全文数据库（硕士） 农业科技辑》 * |
| 王海平等: "离体保存技术在无性繁殖蔬菜种质资源保存中的应用", 《植物遗传资源学报》 * |
| 雷伟侠等: "甘蓝型油菜小孢子培养试管苗的保存和壮苗技术", 《中国油料作物学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Vidal et al. | A temporary immersion system for micropropagation of axillary shoots of hybrid chestnut | |
| CN102668979B (en) | Rooting culture method for poplar tissue culture seedlings | |
| Vijayan et al. | Application of tissue culture techniques for propagation and crop improvement in mulberry (Morus spp.) | |
| CN104099287A (en) | Acquiring and subculture maintaining method for high yield oligomeric proanthocyanidins vitis davidii callus | |
| CN105010147A (en) | Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method | |
| CN101637125B (en) | Propagation method of Chinese yam exsomatize | |
| CN104304238B (en) | The Embedding drying cryopreservation method of downy grape stem apex | |
| CN102405830B (en) | Induction method of rosa chinensis receptacle callus tissues | |
| CN105875413A (en) | Universal induction method for loose type embryonic calluses of beans | |
| CN103798142B (en) | A kind of take stamen as the method that explant sets up lily embryo callus subculture regenerating system | |
| CN106818714A (en) | The cryopreservation method of cowberry plant germplasm | |
| CN103168687B (en) | Method for inducing fast rooting of cluster buds of tobacco leaves | |
| Mukami et al. | Plant regeneration from leaf mesophyll derived protoplasts of cassava (Manihot esculenta Crantz) | |
| CN107950392A (en) | A kind of method for extending Chinese cabbage microspore embryoid regeneration plant subculture interval time | |
| Houllou et al. | Clonal propagation of neem (Azadirachta indica A. Juss.) via direct and indirect in vitro regeneration | |
| Maheshwari et al. | Efficient shoot regeneration from internodal explants of Populus angustifolia, Populus balsamifera and Populus deltoids | |
| CN110923264A (en) | Method suitable for high-efficiency tissue culture of various gramineous plants | |
| CN114793896B (en) | Method for obtaining regenerated complete plants by inducing in-vitro leaves of American pears | |
| CN106613939A (en) | Culture medium for indirect regeneration of high-bush blueberry somatic embryos and culture method | |
| Hembrom et al. | Rapid in vitro production of true-to-type plants of Pogostemon heyneaus through dedieferentiated axillary buds | |
| Zhou et al. | Comparative study of symmetric and asymmetric somatic hybridization between common wheat and Haynaldia villosa | |
| CN205727272U (en) | A kind of stacked seedling cultivation of rice device | |
| CN104604676A (en) | Culture media for tissue culture of red lily | |
| CN103250641A (en) | Devitrification method for vitrified regeneration seedlings of cabbage type rape | |
| Lioshina et al. | Plant regeneration from hairy roots and calluses of periwinkle Vinca minor L. and foxglove purple Digitalis purpurea L. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180424 |