CN106613951A - Method for disinfection of explant in tissue culture - Google Patents
Method for disinfection of explant in tissue culture Download PDFInfo
- Publication number
- CN106613951A CN106613951A CN201610916689.1A CN201610916689A CN106613951A CN 106613951 A CN106613951 A CN 106613951A CN 201610916689 A CN201610916689 A CN 201610916689A CN 106613951 A CN106613951 A CN 106613951A
- Authority
- CN
- China
- Prior art keywords
- explant
- pretreatment
- conditions
- ultrasonic oscillation
- under
- 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
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Apparatus For Disinfection Or Sterilisation (AREA)
Abstract
The embodiment of the invention discloses a method for disinfection of an explant in tissue culture. The method comprises the steps of cleaning the surface of a clean explants, and obtaining the cleaned explant; soaking the cleaned explant in 70% alcohol, performing ultrasonic vibration for a first preset time at a first set temperature, and obtaining a first pretreated explant; soaking the first pretreated explant in 5%-10% NaClO solution, performing ultrasonic vibration for a second preset time at the first set temperature, and obtaining a second pretreated explant; soaking the pretreated explant in sterile water, performing ultrasonic vibration for a second preset time at the first set temperature, and obtaining a third pretreated explant; washing the third pretreated explant, and obtaining a disinfected explant. According to the method for disinfection of the explant in tissue culture, the technical goal of improving cleaning effects and reducing damage rate is achieved.
Description
Technical field
The present invention relates to technical field of tissue culture, more particularly, it relates to explant sterilization in a kind of tissue cultures
Method.
Background technology
The explant material major part of Plant Tissue Breeding takes from field, but because adheres to substantial amounts of on plant surface
Microorganism.Therefore disinfection is had to before vegetable material inoculation, it is therefore an objective to both required various on material surface
Microorganism killing, while and can not damage or only slight damage vegetable material and be unlikely to affect its growth.Therefore, explant
Disinfect be Plant Tissue Breeding work in an important ring.
Problem following prominent is primarily present in the explant disinfecting process commonly used in tissue cultures at present：Explant process
It is improper, cause explant survival rate low or explant sterilization not thoroughly, cause pollution rate high, have impact on tissue culture propagation efficiency and
Yield.
The content of the invention
In view of this, the present invention provides explant sterilization method in a kind of tissue cultures, and the sterilization method disappears to explant
It is malicious thorough, and damage ratio reduction.
A kind of method of explant sterilization in tissue cultures, including：
Explant surface is cleaned up, explant after being cleaned；
Explant after the cleaning is immersed in 70% alcohol, ultrasonic oscillation under the conditions of the first design temperature
One Preset Time, obtains the first pretreatment explant；
Described first pretreatment explant is immersed in the NaClO solution of 5%-10%, in first design temperature
Under the conditions of the Preset Time of ultrasonic oscillation second, obtain the second pretreatment explant；
Described second pretreatment explant is immersed in sterilized water, the ultrasonic wave shake under the conditions of first design temperature
The second Preset Time is swung, the 3rd pretreatment explant is obtained；
Rinse the 3rd pretreatment explant, explant after being sterilized.
Preferably, clean up explant surface to be embodied as：Explant is rinsed 5-7 time with running water, then will be described outer
Implant material is cut into chunks or block or sheet, and running water cleaning explant surface reaches 10-15min.
Preferably, the Preset Time of ultrasonic oscillation first is embodied as under the conditions of the first design temperature：
40 DEG C are warming up to, and under the conditions of 40 DEG C, while using the ultrasonic oscillation 10-20s of 120W.
Preferably, the Preset Time of ultrasonic oscillation second is embodied as under the conditions of first design temperature：
40 DEG C are warming up to, and under the conditions of 40 DEG C, while using the ultrasonic oscillation 5-10min of 120W.
Preferably, rinsing the 3rd pretreatment explant includes：Using the 3rd pretreatment explant described in aseptic water washing
Body material 10-15min.
It can be seen from above-mentioned technical scheme that, explant sterilization method rationally makes in the tissue cultures of the embodiment of the present invention
With lotion, and coordinate the technology of cleaning/vibration, a series of sterilizing operations are carried out to explant, due to time, step and cleaning agent
The correct selection of proportioning, explant cleaning effect is significantly and explant survival rate is high, has reached raising cleaning effect and has reduced damaging
Hinder the technical purpose of rate.
Specific embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described enforcement
Example is only a part of embodiment of the invention, rather than the embodiment of whole.Based on the embodiment in the present invention, this area is common
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belongs to the model of present invention protection
Enclose.
Explant sterilization method in a kind of tissue cultures, it is thorough to explant sterilization to realize, and the skill that damage ratio is reduced
Art purpose.
The method of explant sterilization in the tissue cultures, including：
S11:Explant surface is cleaned up, explant after being cleaned；
S12:Explant after the cleaning is immersed in 70% alcohol, the ultrasonic wave shake under the conditions of the first design temperature
The first Preset Time is swung, the first pretreatment explant is obtained；
S13:Described first pretreatment explant is immersed in the NaClO solution of 5%-10%, in the described first setting
The Preset Time of ultrasonic oscillation second under temperature conditionss, obtains the second pretreatment explant；
S14:Described second pretreatment explant is immersed in sterilized water, the ultrasound under the conditions of first design temperature
Ripple shakes the second Preset Time, obtains the 3rd pretreatment explant；
S15:Rinse the 3rd pretreatment explant, explant after being sterilized.
Above-mentioned sterilization scheme, in order to sterilize good result is reached, comprising multiple sterilisation steps, and using different cleanings
Agent, to ensure to eliminate the purpose of different microorganisms.Meanwhile, in order to not affect explant survival rate difference cleaning agent using suitable
Proportioning is ensureing explant plant safety.
It is as follows：It is the preferred version for above step, it should be noted that：Following scheme is preferably, experimenter
Can in the reasonable scope be selected according to actual conditions, be not limited only to this：
In S11：Clean up explant surface to be embodied as：
Explant is rinsed 5-7 time with running water, then the explant material is cut into chunks or block or sheet, running water is clear
Wash explant surface and reach 10-15min.
Explant is rinsed 5-7 time with running water, with impurity such as the dusts on clean surface.
In S12：The Preset Time of ultrasonic oscillation first is embodied as under the conditions of the first design temperature：
40 DEG C are warming up to, and under the conditions of 40 DEG C, while using the ultrasonic oscillation 10-20s of 120W.
In S13-14：The Preset Time of ultrasonic oscillation second is embodied as under the conditions of first design temperature：
40 DEG C are warming up to, and under the conditions of 40 DEG C, while using the ultrasonic oscillation 5-10min of 120W.
In S15：Rinsing the 3rd pretreatment explant includes：Using the 3rd pretreatment explant described in aseptic water washing
Material 10-15min.
In sum：
Explant sterilization method reasonable employment lotion in the tissue cultures of the embodiment of the present invention, and coordinate cleaning/vibration
Technology, to explant a series of sterilizing operations are carried out, and due to the correct selection of time, step and cleaning agent proportioning, explant is clear
Clean effect is significant and explant survival rate height, have reached the technical purpose for improving cleaning effect and reducing damage ratio.
The foregoing description of the disclosed embodiments, enables professional and technical personnel in the field to realize or using the present invention.
Various modifications to these embodiments will be apparent for those skilled in the art, as defined herein
General Principle can in other embodiments be realized in the case of the spirit or scope without departing from the embodiment of the present invention.Therefore,
The embodiment of the present invention is not intended to be limited to the embodiments shown herein, and be to fit to principles disclosed herein and
The consistent most wide scope of features of novelty.
Claims (5)
1. a kind of method that explant is sterilized in tissue cultures, it is characterised in that include：
Explant surface is cleaned up, explant after being cleaned；
Explant after the cleaning is immersed in 70% alcohol, ultrasonic oscillation first is pre- under the conditions of the first design temperature
If the time, the first pretreatment explant is obtained；
Described first pretreatment explant is immersed in the NaClO solution of 5%-10%, in the first design temperature condition
The lower Preset Time of ultrasonic oscillation second, obtains the second pretreatment explant；
Described second pretreatment explant is immersed in sterilized water, the ultrasonic oscillation the under the conditions of first design temperature
Two Preset Times, obtain the 3rd pretreatment explant；
Rinse the 3rd pretreatment explant, explant after being sterilized.
2. the method that explant is sterilized in tissue cultures as claimed in claim 1, it is characterised in that clean up explant body surface
Face is embodied as：Explant is rinsed 5-7 time with running water, then the explant material is cut into chunks or block or sheet, running water
Cleaning explant surface reaches 10-15min.
3. the method that explant is sterilized in tissue cultures as claimed in claim 1, it is characterised in that
The Preset Time of ultrasonic oscillation first is embodied as under the conditions of the first design temperature：
40 DEG C are warming up to, and under the conditions of 40 DEG C, while using the ultrasonic oscillation 10-20s of 120W.
4. the method that explant is sterilized in tissue cultures as claimed in claim 1, it is characterised in that in the described first setting temperature
The Preset Time of ultrasonic oscillation second is embodied as under the conditions of degree：
40 DEG C are warming up to, and under the conditions of 40 DEG C, while using the ultrasonic oscillation 5-10min of 120W.
5. the method that explant is sterilized in tissue cultures as claimed in claim 1, it is characterised in that rinse the described 3rd pre- place
Reason explant includes：Using the 3rd pretreatment explant material 10-15min described in aseptic water washing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610916689.1A CN106613951A (en) | 2016-10-21 | 2016-10-21 | Method for disinfection of explant in tissue culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610916689.1A CN106613951A (en) | 2016-10-21 | 2016-10-21 | Method for disinfection of explant in tissue culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106613951A true CN106613951A (en) | 2017-05-10 |
Family
ID=58856304
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610916689.1A Pending CN106613951A (en) | 2016-10-21 | 2016-10-21 | Method for disinfection of explant in tissue culture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106613951A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107711516A (en) * | 2017-11-27 | 2018-02-23 | 佛山市恒爱网络科技有限公司 | A kind of method that explant sterilizes in iris tissue cultures |
| CN109588315A (en) * | 2018-12-31 | 2019-04-09 | 西南民族大学 | A kind of sterilization method of bryophyte explant |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102326514A (en) * | 2011-11-01 | 2012-01-25 | 四川中测科技投资有限公司 | Tea tree explant mercury-free disinfectant and explant disinfection method |
| CN103651144A (en) * | 2013-12-20 | 2014-03-26 | 常熟理工学院 | Rapid waxberry propagation method |
| CN104082140A (en) * | 2014-06-30 | 2014-10-08 | 苏州派腾生物医药科技有限公司 | Rapid propagation method of lobed actinostemma herb |
| CN104106464A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
| CN104106465A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
-
2016
- 2016-10-21 CN CN201610916689.1A patent/CN106613951A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102326514A (en) * | 2011-11-01 | 2012-01-25 | 四川中测科技投资有限公司 | Tea tree explant mercury-free disinfectant and explant disinfection method |
| CN103651144A (en) * | 2013-12-20 | 2014-03-26 | 常熟理工学院 | Rapid waxberry propagation method |
| CN104106464A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
| CN104106465A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
| CN104082140A (en) * | 2014-06-30 | 2014-10-08 | 苏州派腾生物医药科技有限公司 | Rapid propagation method of lobed actinostemma herb |
Non-Patent Citations (1)
| Title |
|---|
| 郭勇等: "《植物细胞培养技术与应用》", 31 January 2004 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107711516A (en) * | 2017-11-27 | 2018-02-23 | 佛山市恒爱网络科技有限公司 | A kind of method that explant sterilizes in iris tissue cultures |
| CN109588315A (en) * | 2018-12-31 | 2019-04-09 | 西南民族大学 | A kind of sterilization method of bryophyte explant |
| CN109588315B (en) * | 2018-12-31 | 2021-05-28 | 西南民族大学 | Method for disinfecting bryophyte explants |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102010802A (en) | Concentrated multienzyme cleaning solution and preparation method thereof | |
| CN106613951A (en) | Method for disinfection of explant in tissue culture | |
| CN104106465A (en) | Explant disinfection method in plant tissue culture | |
| CN103418001A (en) | Method for sterilizing animal tissue material and corresponding animal tissue soak solution | |
| CN102934611B (en) | Method for collecting embryo from paeonia suffruticosa seed and for inoculation | |
| CN105830930A (en) | Detoxification and rapid propagation method for edible lily | |
| CN104106464B (en) | The method of explant sterilization in a kind of Plant Tissue Breeding | |
| CN101006790A (en) | Refined iodine disinfectant liquor | |
| CN111265718A (en) | Preparation method of dermal matrix with cell removal and sterilization functions | |
| CN212940634U (en) | Disinfection cabinet for medical operation | |
| US6506417B1 (en) | Composition and process for reducing bacterial citrus canker organisms | |
| CN210936154U (en) | Scalpel cleaning and disinfecting device for otology | |
| CN107711516A (en) | A kind of method that explant sterilizes in iris tissue cultures | |
| CN114653654A (en) | Medical science nursing is with degassing unit that disinfects | |
| CN106171997A (en) | Outer planting surface and the method for endophyte during a kind of plant tissue culture of removal simultaneously | |
| CN104106466A (en) | Explant disinfection method in plant tissue culture | |
| CN217256434U (en) | Clinical laboratory is with disinfection isolation protector that disinfects | |
| CN104115744A (en) | Method for disinfecting explants in plant tissue culture | |
| CN211024280U (en) | Surgical instrument disinfection device for orthopedics department | |
| CN212855854U (en) | Monoclonal antibody re-screening platform | |
| CN103392691A (en) | Container and method for processing, sterilizing and saving biological tissues | |
| CN104115745B (en) | A kind of method of explant sterilization in Plant Tissue Breeding | |
| CN210630587U (en) | Non-thermal physical sterilization device for fresh-cut fruits | |
| CN106568766A (en) | Method for detecting disinfection liquid residue on explant surface during tissue culture | |
| CN214679329U (en) | Paediatrics nursing apparatus places case |
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 |
Application publication date: 20170510 |
|
| RJ01 | Rejection of invention patent application after publication |