CN108260526A - A kind of method for tissue culture of caducous Mazus japonicus - Google Patents
A kind of method for tissue culture of caducous Mazus japonicus Download PDFInfo
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- CN108260526A CN108260526A CN201711267707.9A CN201711267707A CN108260526A CN 108260526 A CN108260526 A CN 108260526A CN 201711267707 A CN201711267707 A CN 201711267707A CN 108260526 A CN108260526 A CN 108260526A
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- culture
- callus
- tissue
- adventitious bud
- caducous
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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
- A01H4/001—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
- A01H4/008—Methods for regeneration to complete plants
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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)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to field of plant tissue culture, disclose a kind of method for tissue culture of caducous Mazus japonicus, include the following steps：1) blade forms callus：By the caducous Mazus japonicus blade of field picking after disinfecting, vein and leaf margin are cut, the fritter of about 5mm × 5mm is cut into, is seeded on callus inducing medium, routine culture is to forming callus；2) callus differentiation adventitious bud：The callus formed in step 1) is cut into the fritter of about 5mm × 5mm, is seeded on the inducing culture of Bud Differentiation, adventitious bud is formed on routine culture to callus；3) adventitious bud rooting：The adventitious bud of about 3 5cm of size formed in step 2) is cut, is seeded in rooting induction culture medium, routine culture to root is formed；4) tissue-cultured seedling is bloomed.The method of the present invention can obtain a large amount of excellent caducous Mazus japonicus tissue-cultured seedling within 50d, and market application foreground is wide.
Description
Technical field
The present invention relates to field of plant tissue culture, specifically, being related to a kind of method for tissue culture of caducous Mazus japonicus.
Background technology
Caducous Mazus japonicus (Mazus caducifer Hance) belongs to Scrophulariaceae Mazus perennial herb, high 20-
50cm, it is sturdy, it is all by the long pubescence of many cells white；Raceme basidixed is long up to 35cm, the month at florescence 4-5；Capsule ball
Shape, the fruiting period 6-8 months；Seed sepia is more and small；It is grown on dark and damp roadside below height above sea level 1300m, hayashishita, Cao Po.Logical spring
Careless platymiscium is short sward sheet, mostly several to have medical value, has the clearing heat and detoxicating, functions such as crazy poison, analgesic, stomach invigorating of dispelling, controls partially
The diseases such as headache, furunculosis, running sore and scald, nameless gall.The platymiscium resource is relatively abundant, and wild shape is rendered as in most area
State need the excellent type of therefrom selection traits and applies in practice to come, such as medicinal, landscape application etc..Have no early at present
The relevant document of Mazus japonicus tissue cultures and patent are fallen, it can be to the excellent caducous Mazus japonicus product of selection using tissue culture technique
Kind carries out rapid, high volume breeding, and keeps maternal fine quality, and the application for caducous Mazus japonicus provides a large amount of excellent former materials
Material.
Invention content
To solve at present the caducous Mazus japonicus in wild state mostly, improved seeds are selected, using tissue culture technique
A large amount of quick breedings are carried out to it, a large amount of excellent caducous Mazus japonicus raw material are provided, are worth for its medicinal and landscape application
To extensively using laying the foundation, the present invention provides a kind of method for tissue culture of caducous Mazus japonicus.
A kind of method for tissue culture of caducous Mazus japonicus provided by the invention, includes the following steps：
1) blade forms callus：By the caducous Mazus japonicus blade of field picking after disinfecting, vein is cut
And leaf margin, the fritter of about 5mm × 5mm is cut into, is seeded on callus inducing medium, routine culture to formation callus group
It knits；
2) callus differentiation adventitious bud：The callus formed in step 1) is cut into the fritter of about 5mm × 5mm, is connect
On kind to the inducing culture of Bud Differentiation, adventitious bud is formed on routine culture to callus；
3) adventitious bud rooting：The adventitious bud of the size about 3-5cm formed in step 2) is cut, is seeded to rooting induction training
It supports in base, routine culture to root is formed；
4) tissue-cultured seedling is bloomed：The tissue-cultured seedling taken root continuation in root media is subjected to routine culture, petal can be formed,
Until blooming.
Wherein, the blade forms the inducing culture of callus to include 1mg/L 6-benzyl aminopurines and 0.3mg/
The MS culture mediums of L methyl α-naphthyl acetates.
Wherein, the inducing culture that the callus differentiates adventitious bud be comprising 2mg/L 6-benzyl aminopurines and
The MS culture mediums of 0.1mg/L methyl α-naphthyl acetates.
Wherein, the inducing culture of the adventitious bud rooting is includes 1mg/L 6-benzyl aminopurines and 0.3mg/L naphthalene second
The MS culture mediums of acid.
Wherein, the inducing culture that the tissue-cultured seedling is bloomed is includes 1mg/L 6-benzyl aminopurines and 0.3mg/L naphthalene second
The MS culture mediums of acid.
Wherein, it is 30g/L, a concentration of 8g/L of agar powder containing sucrose concentration in the MS culture mediums.
Wherein, it is 6.0 that the condition of culture of the routine culture, which is pH value, intensity of illumination 1500lx, light application time 10h/d,
Constant incubator temperature is 25 DEG C.
Wherein, the time that the blade forms callus is 20d.
Wherein, the time that the callus differentiates adventitious bud is 10d.
Wherein, the time of the adventitious bud rooting is 20d.
Wherein, the time that the tissue-cultured seedling forms petal is 10d.
The beneficial effects of the present invention are：
1) method of the invention can obtain a large amount of excellent caducous Mazus japonicus tissue-cultured seedling within 50d, can quickly, great Liang He
The good application for caducous Mazus japonicus provides a large amount of excellent raw material.
2) method of the invention can obtain the caducous Mazus japonicus tissue-cultured seedling that a large amount of excellent confessions watch, market within 60d
It has a extensive future.
3) method of the invention has in protection wild caducous Mazus japonicus natural resources and good ecological environment etc.
Significance.
Description of the drawings
Fig. 1：It is callus (the caducous Mazus japonicus that caducous Mazus japonicus blade is formed after cultivation 20d in the embodiment of the present invention
It is picked from field, similarly hereinafter).
Fig. 2：The adventitious bud differentiated after the callus tissue culture 10d for being caducous Mazus japonicus in the embodiment of the present invention.
Fig. 3：It is the situation of taking root in the embodiment of the present invention after caducous Mazus japonicus Adventitious bud culture 20d.
Fig. 4：It is the seedling of blooming that caducous Mazus japonicus tissue-cultured seedling cultivates after 10d in the embodiment of the present invention.
Specific embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It is it will be appreciated that following real
Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this field
Art personnel can carry out various modifications and replace to the present invention in the case of without departing substantially from spirit of the invention and spirit.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1
A kind of method for tissue culture of caducous Mazus japonicus, includes the following steps：
1) blade forms callus：By the caducous Mazus japonicus blade of field picking after disinfecting, vein is cut
And leaf margin, the fritter of about 5mm × 5mm is cut into, is seeded on callus inducing medium, routine culture to formation callus group
It knits；The inducing culture that the blade forms callus is comprising 1mg/L 6-benzyl aminopurines and 0.3mg/L methyl α-naphthyl acetates
MS culture mediums；The time that the blade forms callus is 20d；Contain sucrose concentration in the MS culture mediums for 30g/L, fine jade
A concentration of 8g/L of cosmetics；The condition of culture of the routine culture is that pH value is 6.0, intensity of illumination 1500lx, light application time 10h/
D, constant incubator temperature are 25 DEG C.
2) callus differentiation adventitious bud：The callus formed in step 1) is cut into the fritter of about 5mm × 5mm, is connect
On kind to the inducing culture of Bud Differentiation, adventitious bud is formed on routine culture to callus；The callus differentiates not
The inducing culture of normal bud is the MS culture mediums comprising 2mg/L 6-benzyl aminopurines and 0.1mg/L methyl α-naphthyl acetates, and callus divides
The time for dissolving adventitious bud is 10d；The same step 1) of condition of culture of the MS culture mediums and routine culture.
3) adventitious bud rooting：The adventitious bud of the size about 3-5cm formed in step 2) is cut, is seeded to rooting induction training
It supports in base, routine culture to root is formed；The inducing culture of the adventitious bud rooting be comprising 1mg/L 6-benzyl aminopurines and
The MS culture mediums of 0.3mg/L methyl α-naphthyl acetates；The time that the callus differentiates adventitious bud is 20d；The MS culture mediums
With the same step 1) of condition of culture of routine culture.
4) tissue-cultured seedling is bloomed：The tissue-cultured seedling taken root continuation in root media is subjected to routine culture, petal can be formed,
Until blooming.The inducing culture that the tissue-cultured seedling is bloomed is comprising 1mg/L 6-benzyl aminopurines and 0.3mg/L methyl α-naphthyl acetates
MS culture mediums；The time that the tissue-cultured seedling forms petal is 10d；The MS culture mediums and the condition of culture of routine culture are same
Step 1).
The result shows that method of the invention can obtain a large amount of excellent caducous Mazus japonicus tissue-cultured seedling within 50d, it can be fast
Fast, a large amount of and good application for caducous Mazus japonicus provides a large amount of excellent raw material；The method of the present invention can within 60d
The caducous Mazus japonicus tissue-cultured seedling that a large amount of excellent confessions watch is obtained, market application foreground is wide.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. a kind of method for tissue culture of caducous Mazus japonicus, includes the following steps：
1) blade forms callus：By the caducous Mazus japonicus blade of field picking after disinfecting, vein and leaf are cut
Edge is cut into the fritter of about 5mm × 5mm, is seeded on callus inducing medium, and routine culture is to forming callus；
2) callus differentiation adventitious bud：The callus formed in step 1) is cut into the fritter of about 5mm × 5mm, is seeded to
On the inducing culture of Bud Differentiation, adventitious bud is formed on routine culture to callus；
3) adventitious bud rooting：The adventitious bud of the size about 3-5cm formed in step 2) is cut, is seeded to rooting induction culture medium
In, routine culture to root is formed；
4) tissue-cultured seedling is bloomed：The tissue-cultured seedling taken root continuation in root media is subjected to routine culture, petal can be formed, until
It blooms.
2. the method as described in claim 1, which is characterized in that the inducing culture that the blade forms callus be comprising
The MS culture mediums of 1mg/L 6-benzyl aminopurines and 0.3mg/L methyl α-naphthyl acetates.
3. the method as described in claim 1, which is characterized in that the inducing culture that the callus differentiates adventitious bud is
MS culture mediums comprising 2mg/L 6-benzyl aminopurines and 0.1mg/L methyl α-naphthyl acetates.
4. the method as described in claim 1, which is characterized in that the inducing culture of the adventitious bud rooting is includes 1mg/L
The MS culture mediums of 6-benzyl aminopurine and 0.3mg/L methyl α-naphthyl acetates.
5. the method as described in claim 1, which is characterized in that the inducing culture that the tissue-cultured seedling is bloomed is comprising 1mg/L
The MS culture mediums of 6-benzyl aminopurine and 0.3mg/L methyl α-naphthyl acetates.
6. the method as described in claim 1, which is characterized in that the time that the blade forms callus is 20d.
7. the method as described in claim 1, which is characterized in that the time that the callus differentiates adventitious bud is 10d.
8. the method as described in claim 1, which is characterized in that the time of the adventitious bud rooting is 20d.
9. the method as described in claim 1, which is characterized in that the time that the tissue-cultured seedling forms petal is 10d.
10. claim 1-9 any one of them methods are the applications in a kind of method for tissue culture of caducous Mazus japonicus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711267707.9A CN108260526B (en) | 2017-12-05 | 2017-12-05 | Tissue culture method of early-falling marsdenia tenacissima |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711267707.9A CN108260526B (en) | 2017-12-05 | 2017-12-05 | Tissue culture method of early-falling marsdenia tenacissima |
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| Publication Number | Publication Date |
|---|---|
| CN108260526A true CN108260526A (en) | 2018-07-10 |
| CN108260526B CN108260526B (en) | 2021-01-12 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711267707.9A Active CN108260526B (en) | 2017-12-05 | 2017-12-05 | Tissue culture method of early-falling marsdenia tenacissima |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103704135A (en) * | 2013-12-20 | 2014-04-09 | 长沙学院 | In-vitro rapid propagation method for plantains |
| CN105766630A (en) * | 2014-12-19 | 2016-07-20 | 岳兰兰 | Radix rehmanniae virus free tissue culture rapid propagation method |
-
2017
- 2017-12-05 CN CN201711267707.9A patent/CN108260526B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103704135A (en) * | 2013-12-20 | 2014-04-09 | 长沙学院 | In-vitro rapid propagation method for plantains |
| CN105766630A (en) * | 2014-12-19 | 2016-07-20 | 岳兰兰 | Radix rehmanniae virus free tissue culture rapid propagation method |
Non-Patent Citations (2)
| Title |
|---|
| 宾金华等: "蓝猪耳再生系统的建立", 《广西农业生物科学》 * |
| 陈敏艳: "地黄组织培养的研究", 《中国优秀硕士学位论文全文数据库》 * |
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| CN108260526B (en) | 2021-01-12 |
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