CN108834892A - A kind of Chinese herbaceous peony callus induction method - Google Patents
A kind of Chinese herbaceous peony callus induction method Download PDFInfo
- Publication number
- CN108834892A CN108834892A CN201810672767.7A CN201810672767A CN108834892A CN 108834892 A CN108834892 A CN 108834892A CN 201810672767 A CN201810672767 A CN 201810672767A CN 108834892 A CN108834892 A CN 108834892A
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- CN
- China
- Prior art keywords
- herbaceous peony
- chinese herbaceous
- callus induction
- induction method
- culture medium
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- 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.)
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Classifications
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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
Abstract
The invention discloses a kind of Chinese herbaceous peony callus induction methods, in mid-March to mid-April, the tender explant of clip Chinese herbaceous peony children, rinses 2h with flowing water, is placed in 2%NaClO solution and sterilizes 20min, it is cleaned with sterile water, it is put into 70% alcohol and sterilizes, after aseptic water washing is clean, blot explant surface moisture, it is seeded in MS culture medium, culturing room's condition is：22 DEG C of temperature, dark treatment.Also containing substances such as 6-BA, NAA, 2,4-D in MS culture medium.Chinese herbaceous peony callus differentiation after this method is establishes good basis with the domestication of plant, and easy to operate, low in cost, has good market prospects, can provide good help for the breeding and growth of Chinese herbaceous peony.
Description
Technical field
The present invention relates to a kind of Chinese herbaceous peony difference explant callus induction methods.
Background technique
Chinese herbaceous peony (Paeonia lactiflora Pall.) is that Paeoniaceae (Paeoniaceae) Paeonia (Paeonia) is more
Year life persistent root herbs, are distributed in Jiangsu, northeast, one band of North China, Shaanxi and SOUTH OF GANSU in China.Chinese herbaceous peony is the biography in China
System famous flower is described as " Hua Xian " and " flower phase " by people, and be otherwise known as " flora in May ".
For the propagation method of Chinese herbaceous peony mainly based on offshoot, this traditional propagation method has period length, low efficiency, numerous
It grows the disadvantages such as coefficient is low and is difficult meet the needs of market still to form sound industry production mechanism.And in Chinese herbaceous peony tissue cultures
There are still the browning death rate it is high, contamination phenomenon is serious the problems such as hinder the progress of its research and development.
Summary of the invention
The object of the present invention is to provide a kind of Chinese herbaceous peony difference explant callus induction methods.
The purpose of the present invention is what is be achieved through the following technical solutions：
Chinese herbaceous peony callus induction method of the invention, including step：
In mid-March to mid-April, the tender explant of clip Chinese herbaceous peony children is packed into gauze on beaker cover after cleaning, is rushed with flowing water
It washes, is placed in NaClO solution and sterilizes, cleaned with sterile water, be put into alcohol and sterilize, after aseptic water washing is clean, blot explant
Body surface moisture is seeded in MS culture medium, and culturing room's condition is：22 DEG C of temperature, dark treatment.
As seen from the above technical solution provided by the invention, the present invention implements the Chinese herbaceous peony callus induction provided
Good basis is established in the domestication of method, the differentiation of Chinese herbaceous peony callus and plant after being, and easy to operate, low in cost, is had
Good market prospects.Good help can be provided for the breeding and growth of Chinese herbaceous peony.
Specific embodiment
The embodiment of the present invention will be described in further detail below.What is be not described in detail in the embodiment of the present invention is interior
Appearance belongs to the prior art well known to professional and technical personnel in the field.
Chinese herbaceous peony callus induction method of the invention, preferable specific embodiment are：
Including step：
In mid-March to mid-April, the tender explant of clip Chinese herbaceous peony children is packed into gauze on beaker cover after cleaning, is rushed with flowing water
It washes, is placed in NaClO solution and sterilizes, cleaned with sterile water, be put into alcohol and sterilize, after aseptic water washing is clean, blot explant
Body surface moisture is seeded in MS culture medium, and culturing room's condition is：22 DEG C of temperature, dark treatment.Above-mentioned flowing water washing time is
2h。
Above-mentioned disinfection NaClO concentration is 2%, disinfecting time 20min.
Above-mentioned rubbing alcohol concentration is 70%, disinfecting time 8-10s.
Above-mentioned is twice 3-5 times with aseptic water washing number.
Also contain following any one or more adding ingredients in the MS culture medium：The substances such as 6-BA, NAA, 2,4-D.
Stem section is MS+6-BA 1mg/L+2,4-D 2.0mg/L with the MS culture medium adding ingredient.
Blade is MS+6-BA 1mg/L+NAA 1.0mg/L with the MS culture medium adding ingredient.
Anther is MS+6-BA 0.5mg/L+2,4-D 1.5mg/L with the MS culture medium adding ingredient.
Petal is MS+6-BA 0.5mg/L+2,4-D 2.0mg/L with the MS culture medium adding ingredient
The domestication of the method for Chinese herbaceous peony callus induction of the invention, the differentiation of Chinese herbaceous peony callus and plant after being is established
Determine good basis, and easy to operate, low in cost, there are good market prospects, can lay the foundation for Chinese herbaceous peony tissue cultures
Specific implementation：
It draws materials the time：In mid or late March, 2018 is to early or mid April.
Selection：The tender explant of Chinese herbaceous peony children.
Explant sterilizing and inoculation：
The tender explant of Chinese herbaceous peony is rinsed well with clear water, then is handled with flowing water 2h, the immersion that concentration is 5% is placed in,
Disinfecting time is 20min.
The Chinese herbaceous peony explant being soaked in 2%NaClO solution is put into super-clean bench, for aseptic water washing 3-5 after, use is dense
Degree impregnates 8-10s for 70% alcohol, rear to use aseptic water washing 3-5 times.
Explant is placed in suck dry moisture on aseptic filter paper, and stem section cuts marginal portion and intermediate to cutting, blade and petal
Marginal portion is cut, expands the contact area of explant and culture medium as far as possible, is inoculated on culture medium, pays attention to avoiding intersecting feeling
Dye, Primary culture.
Callus induction：
Culture medium prescription：It is preferred that stem section culture base is MS+6-BA 1mg/L+2,4-D 2mg/L, leaf culture base is MS+
6-BA 1mg/L+NAA 1mg/L, anther and petal swirl burner base are MS+6-BA 0.5mg/L+2,4-D1.5mg/L, pH 5.8.
Condition of culture：22 DEG C of temperature, dark treatment.
As a result with analysis：
The substances such as 6-BA, NAA, 2, the 4-D of independent assortment various concentration and each combined concentration are freely inoculated with appropriate
Chinese herbaceous peony explant body portion, stem section, blade, anther, petal.
Influence of the 1 different culture medium formula of table to different explant inductivities
Test result shows (table 1), and according to inductivity, preferably stem section MS culture medium is MS+6-BA 1mg/L+NAA2mg/
L, blade MS culture medium are MS+6-BA 1mg/L+NAA 1mg/L, and anther culture medium is MS+6-BA 0.5mg/L+2,4-D
1.5mg/L, petal swirl burner base MS+6-BA 0.5mg/L+2,4-D 2.0mg/L.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Within the technical scope of the present disclosure, any changes or substitutions that can be easily thought of by anyone skilled in the art,
It should be covered by the protection scope of the present invention.Therefore, protection scope of the present invention should be with the protection model of claims
Subject to enclosing.
Claims (10)
1. a kind of Chinese herbaceous peony callus induction method, which is characterized in that including step：
In mid-March to mid-April, the tender explant of clip Chinese herbaceous peony children is packed into gauze on beaker cover after cleaning, is rinsed with flowing water,
It is placed in NaClO solution and sterilizes, cleaned with sterile water, be put into alcohol and sterilize, after aseptic water washing is clean, blot explant body surface
Face moisture is seeded in MS culture medium, and culturing room's condition is：22 DEG C of temperature, dark treatment.
2. Chinese herbaceous peony callus induction method according to claim 1, which is characterized in that above-mentioned flowing water washing time is 2h.
3. Chinese herbaceous peony callus induction method according to claim 1, which is characterized in that above-mentioned disinfection is with NaClO concentration
2%, disinfecting time 20min.
4. Chinese herbaceous peony callus induction method according to claim 1, which is characterized in that above-mentioned rubbing alcohol concentration is
70%, disinfecting time 8-10s.
5. Chinese herbaceous peony callus induction method according to claim 1, which is characterized in that above-mentioned twice with aseptic water washing
Number is 3-5 times.
6. Chinese herbaceous peony callus induction method according to claim 1, which is characterized in that in the MS culture medium also containing with
Under any one or more adding ingredients：6-BA,NAA,2,4-D.
7. Chinese herbaceous peony callus induction method according to claim 6, which is characterized in that stem section is added with the MS culture medium
Ingredient is MS+6-BA 1mg/L+NAA 2mg/L.
8. Chinese herbaceous peony callus induction method according to claim 6, which is characterized in that blade is added with the MS culture medium
Ingredient is MS+6-BA 1mg/L+NAA 1mg/L.
9. Chinese herbaceous peony callus induction method according to claim 6, which is characterized in that anther is added with the MS culture medium
Ingredient is MS+6-BA 0.5mg/L+2,4-D 1.5mg/L.
10. Chinese herbaceous peony callus induction method according to claim 6, which is characterized in that petal is added with the MS culture medium
Addition is divided into MS+6-BA 0.5mg/L+2,4-D 2.0mg/L.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2018106226601 | 2018-06-15 | ||
| CN201810622660 | 2018-06-15 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108834892A true CN108834892A (en) | 2018-11-20 |
Family
ID=64202386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810672767.7A Pending CN108834892A (en) | 2018-06-15 | 2018-06-26 | A kind of Chinese herbaceous peony callus induction method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113678732A (en) * | 2021-08-17 | 2021-11-23 | 菏泽学院 | Peony stem leaf callus induction method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0523178A (en) * | 1991-07-24 | 1993-02-02 | Mitsubishi Agricult Mach Co Ltd | Tissue culture of tree-peony |
| CN102144563A (en) * | 2009-05-11 | 2011-08-10 | 中国林业科学研究院林业研究所 | Method for inducing calluses of peony anther |
| CN107593446A (en) * | 2017-10-19 | 2018-01-19 | 寿县正阳油用牡丹种植专业合作社 | A kind of fast breeding method of the induction and regeneration of tree peony |
-
2018
- 2018-06-26 CN CN201810672767.7A patent/CN108834892A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0523178A (en) * | 1991-07-24 | 1993-02-02 | Mitsubishi Agricult Mach Co Ltd | Tissue culture of tree-peony |
| CN102144563A (en) * | 2009-05-11 | 2011-08-10 | 中国林业科学研究院林业研究所 | Method for inducing calluses of peony anther |
| CN107593446A (en) * | 2017-10-19 | 2018-01-19 | 寿县正阳油用牡丹种植专业合作社 | A kind of fast breeding method of the induction and regeneration of tree peony |
Non-Patent Citations (2)
| Title |
|---|
| 张玉芳: "牡丹愈伤组织诱导的初步研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 朱向涛: "凤丹体胚发生及愈伤组织诱导芽分化研究", 《中国博士学位论文全文数据库 农业科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113678732A (en) * | 2021-08-17 | 2021-11-23 | 菏泽学院 | Peony stem leaf callus induction method |
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Application publication date: 20181120 |
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