CN104106464B - The method of explant sterilization in a kind of Plant Tissue Breeding - Google Patents
The method of explant sterilization in a kind of Plant Tissue Breeding Download PDFInfo
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- CN104106464B CN104106464B CN201410261359.4A CN201410261359A CN104106464B CN 104106464 B CN104106464 B CN 104106464B CN 201410261359 A CN201410261359 A CN 201410261359A CN 104106464 B CN104106464 B CN 104106464B
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- explant
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Abstract
The invention discloses the method for explant sterilization in a kind of Plant Tissue Breeding, material running water is adopted to clean explant material, then oxirane is adopted to carry out disinfection to the explant material after cleaning, finally adopt thimerosal to carry out disinfection liquid process to explant material down ultrasonic, complete sterilization.The method sterilization is easy, bactericidal effect good, and toxicity is little, not easily attacking material.
Description
Technical field
The present invention relates to field of plant tissue culture technique, particularly relate to the method for explant sterilization in a kind of Plant Tissue Breeding.
Background technology
The explant major part of Plant Tissue Breeding takes from field, adheres to a large amount of microorganisms on the surface, and this is a large obstacle of tissue cultures.Therefore must disinfect before material inoculated and cultured, sterilization requires the various microorganism killings on material surface on the one hand, can not damage again or only slight damage organization material and do not affect its growth simultaneously.Therefore, disinfecting of explant is an important ring in Plant Tissue Breeding work.
Existing sterilization method, Disinfection Effect is bad, and the microorganism on material can not be killed to greatest extent, and existing sterilization method, explant is vulnerable to infringement, greatly reduces the survival rate of Plant Tissue Breeding.
Summary of the invention
The object of this part is some aspects of general introduction embodiments of the invention and briefly introduces some preferred embodiments.May do in the specification digest and denomination of invention of this part and the application a little simplify or omit with avoid making this part, specification digest and denomination of invention object fuzzy, and this simplification or omit and can not be used for limiting the scope of the invention.
In view of Problems existing in explant disinfection method in above-mentioned and/or existing Plant Tissue Breeding, propose the present invention.
Therefore, the present invention wherein object is to provide the method for explant sterilization in a kind of Plant Tissue Breeding, this sterilization method, and its sterilization is easy, Disinfection Effect is good, and toxicity is little, not attacking material.
For solving the problems of the technologies described above, according to an aspect of the present invention, the invention provides following technical scheme: the method for explant sterilization in a kind of Plant Tissue Breeding, material running water is adopted to clean explant material, then oxirane is adopted to carry out disinfection to the explant material after cleaning, finally adopt thimerosal to carry out disinfection liquid process to explant material down ultrasonic, complete sterilization.
As the present invention's preferred embodiment, described thimerosal is the mixed solution of mercuric chloride and tween.
As the present invention's preferred embodiment, it specifically comprises the steps:
Step one: utilize tap water explant material 30 ~ 50min;
Step 2: at room temperature, utilizes oxirane to carry out disinfection 2 ~ 4 hours to the explant material after above-mentioned cleaning, completes pretreatment;
Step 3: above-mentioned pretreated explant material is immersed containing 0.025 ~ 1% mercuric chloride and 0.1 ~ 0.2% tween mixed solution in, and with Vltrasonic device ultrasonic oscillation 5 ~ 15min, then use sterile water wash explant material 5 ~ 7 times.
As the present invention's preferred embodiment, in described step one, the time of tap water explant material is 50min.
As the present invention's preferred embodiment, in described step 2, the time adopting oxirane to carry out disinfection to the explant material after cleaning is 3 hours.
As the present invention's preferred embodiment, in described step 3, pretreated explant material is immersed containing 0.025% mercuric chloride and 0.1% tween mixed solution in, and ultrasonic oscillation 10min.
As the present invention's preferred embodiment, in described step 3, the number of times adopting sterile water wash explant material is 6 times.
As the present invention's preferred embodiment, the power of described Vltrasonic device is 90W.
The invention provides the method for explant sterilization in a kind of Plant Tissue Breeding, its sterilization is easy, bactericidal effect good, and toxicity is little, and explant material is easy damaged not.
Embodiment
For enabling above-mentioned purpose of the present invention, feature and advantage become apparent more, are described in detail the specific embodiment of the present invention below in conjunction with accompanying drawing.
Set forth a lot of detail in the following description so that fully understand the present invention, but the present invention can also adopt other to be different from alternate manner described here to implement, those skilled in the art can when without prejudice to doing similar popularization when intension of the present invention, therefore the present invention is by the restriction of following public specific embodiment.
The invention provides the method for explant sterilization in a kind of Plant Tissue Breeding, material running water is adopted to clean explant material, then oxirane is adopted to carry out disinfection to the explant material after cleaning, finally adopt thimerosal to carry out disinfection liquid process to explant material down ultrasonic, complete sterilization.
Below adopt embodiment to describe embodiments of the present invention in detail, to the present invention, how application technology means solve technical problem whereby, and the implementation procedure reaching technique effect can fully understand and implement according to this.
It should be noted that, the mixed solution that the thimerosal that the present invention uses is mercuric chloride and tween.
embodiment 1
Explant material is cut into segment or fritter or sheet or thread, then carries out disinfection according to the following steps:
Step one: utilize tap water explant material 30min;
Step 2: at room temperature, utilizes oxirane to carry out disinfection 2 hours to the explant material after above-mentioned cleaning, completes pretreatment;
Step 3: above-mentioned pretreated explant material is immersed containing 0.025% mercuric chloride and 0.1% tween mixed solution in, and with Vltrasonic device ultrasonic oscillation 5min, then use sterile water wash explant material 5 times, complete sterilization.
embodiment 2
Explant material is cut into segment or fritter or sheet or thread, then carries out disinfection according to the following steps:
Step one: utilize tap water explant material 50min;
Step 2: at room temperature, utilizes oxirane to carry out disinfection 3 hours to the explant material after above-mentioned cleaning, completes pretreatment;
Step 3: above-mentioned pretreated explant material is immersed containing 0.025% mercuric chloride and 0.1% tween mixed solution in, and with Vltrasonic device ultrasonic oscillation 10min, then use sterile water wash explant material 6 times, complete sterilization.
embodiment 3
Explant material is cut into segment or fritter or sheet or thread, then carries out disinfection according to the following steps:
Step one: utilize tap water explant material 40min;
Step 2: at room temperature, utilizes oxirane to carry out disinfection 4 hours to the explant material after above-mentioned cleaning, completes pretreatment;
Step 3: above-mentioned pretreated explant material is immersed containing 1% mercuric chloride and 0.2% tween mixed solution in, and with Vltrasonic device ultrasonic oscillation 15min, then use sterile water wash explant material 7 times, complete sterilization.
embodiment 4
Explant material is cut into segment or fritter or sheet or thread, then carries out disinfection according to the following steps:
Step one: utilize tap water explant material 50min;
Step 2: at room temperature, utilizes oxirane to carry out disinfection 2 hours to the explant material after above-mentioned cleaning, completes pretreatment;
Step 3: above-mentioned pretreated explant material is immersed containing 0.025% mercuric chloride and 0.1% tween mixed solution in, and with Vltrasonic device ultrasonic oscillation 10min, then use sterile water wash explant material 5 times, complete sterilization.
sterilization effect
Select the sterilization method of the embodiment of the present invention 1 ~ 4 and traditional sterilizing methods to carry out tissue culture experiments to explant, its aseptic rate contrast table is in table 1:
Table 1 sterilization method of the present invention and traditional method for disinfection are to the killing effect of bacterium and fungi
| Sequence number | Sterilization complexity | Aseptic rate (%) |
| Embodiment 1 | Easily | 96 |
| Embodiment 2 | The easiest | 100 |
| Embodiment 3 | The easiest | 100 |
| Embodiment 4 | Easily | 99 |
| Conventional antimicrobial method | Not easily | 20.1 |
It should be noted that, the present invention does not limit the kind of explant and position.
As can be seen here, the method for explant sterilization in Plant Tissue Breeding of the present invention, its sterilization is easy, bactericidal effect good, and toxicity is little, not easy damaged explant material.
It should be noted that, above embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to technical scheme of the present invention or equivalent replacement, and not departing from the spirit and scope of technical solution of the present invention, it all should be encompassed in the middle of right of the present invention.
Claims (1)
1. the method for explant sterilization in a Plant Tissue Breeding, it is characterized in that: adopt running water to clean explant material, then oxirane is adopted to carry out disinfection to the explant material after cleaning, thimerosal is finally adopted to carry out disinfection liquid process to explant material down ultrasonic, complete sterilization
Described thimerosal is the mixed solution of mercuric chloride and tween,
It specifically comprises the steps:
Step one: utilize tap water explant material 30 ~ 50min;
Step 2: at room temperature, utilizes oxirane to carry out disinfection 2 ~ 4 hours to the explant material after above-mentioned cleaning, completes pretreatment;
Step 3: above-mentioned pretreated explant material is immersed containing 0.025 ~ 1% mercuric chloride and 0.1 ~ 0.2% tween mixed solution in, and with Vltrasonic device ultrasonic oscillation 5 ~ 15min, then use sterile water wash explant material 5 ~ 7 times,
The power of described Vltrasonic device is 90W.
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| CN201410261359.4A CN104106464B (en) | 2014-06-12 | 2014-06-12 | The method of explant sterilization in a kind of Plant Tissue Breeding |
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| CN201410261359.4A CN104106464B (en) | 2014-06-12 | 2014-06-12 | The method of explant sterilization in a kind of Plant Tissue Breeding |
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| CN104106464B true CN104106464B (en) | 2016-03-30 |
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| CN105850730A (en) * | 2016-04-06 | 2016-08-17 | 南京晓庄学院 | Plant material surface disinfection method |
| CN106613951A (en) * | 2016-10-21 | 2017-05-10 | 定州市绿谷农业科技发展有限公司 | Method for disinfection of explant in tissue culture |
| CN106888976A (en) * | 2017-04-01 | 2017-06-27 | 中国农业科学院草原研究所 | Plant Tissue Breeding Explant sterilizing device |
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| CN102823497A (en) * | 2012-09-19 | 2012-12-19 | 广东省林业科学研究院 | Clonal tissue culture breeding method of Liquidambar formosana Hance |
| CN103120126A (en) * | 2013-02-04 | 2013-05-29 | 南京工业大学 | Method for carrying out anoectochilus formosanus tissue culture propagation by using intermittent immersion bioreactor |
| CN103125389A (en) * | 2013-03-04 | 2013-06-05 | 北京农业生物技术研究中心 | Method for increasing cycle conversion utilization of Zoysia japonica high frequency regeneration series |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102823497A (en) * | 2012-09-19 | 2012-12-19 | 广东省林业科学研究院 | Clonal tissue culture breeding method of Liquidambar formosana Hance |
| CN103120126A (en) * | 2013-02-04 | 2013-05-29 | 南京工业大学 | Method for carrying out anoectochilus formosanus tissue culture propagation by using intermittent immersion bioreactor |
| CN103125389A (en) * | 2013-03-04 | 2013-06-05 | 北京农业生物技术研究中心 | Method for increasing cycle conversion utilization of Zoysia japonica high frequency regeneration series |
Non-Patent Citations (1)
| Title |
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| 环氧乙烷灭菌;黄靖雄;《中华医院感染学杂志》;20041231;第14卷(第12期);第1435-1439页 * |
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