CN104106465A - Explant disinfection method in plant tissue culture - Google Patents
Explant disinfection method in plant tissue culture Download PDFInfo
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- CN104106465A CN104106465A CN201410262385.9A CN201410262385A CN104106465A CN 104106465 A CN104106465 A CN 104106465A CN 201410262385 A CN201410262385 A CN 201410262385A CN 104106465 A CN104106465 A CN 104106465A
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Abstract
An explant disinfection method in plant tissue culture is disclosed. An explant material is dipped in a potassium permanganate solution, washed by tap water, dipped in a disinfectant and subjected to ultrasonic treatment for a preset period of time at the same time, and finally washed by sterile water for a plurality of times so that disinfection is completed. The method has characteristics of easy disinfection, good disinfection effects, low toxicity, and difficult damaging for the material.
Description
Technical field
The present invention relates to field of plant tissue culture technique, relate in particular to the method for explant sterilization in a kind of Plant Tissue Breeding.
Background technology
The explant major part that Plant Tissue Breeding is used is taken from field, adheres to a large amount of microorganisms on surface, and this is the large obstacle that tissue is cultivated.Therefore before material inoculated and cultured, must disinfect, sterilization requires the various microorganism killings on material surface on the one hand, can not damage again or slight damage organization material and do not affect its growth only simultaneously.Therefore, disinfecting of explant is the 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 to summarize some aspects of embodiments of the invention and briefly introduces some preferred embodiments.In this part and the application's specification digest and denomination of invention, may do a little simplification or omit to avoid the making object of this part, specification digest and denomination of invention fuzzy, and this simplification or omit and can not be used for limiting the scope of the invention.
In view of the problem existing in explant sterilization method in above-mentioned and/or existing Plant Tissue Breeding, the present invention is proposed.
Therefore, the present invention wherein object is to provide a kind of method that in Plant Tissue Breeding, explant is sterilized, this sterilization method, and its sterilization is easy, Disinfection Effect is good, and toxicity is little, does not damage material.
For solving the problems of the technologies described above, according to an aspect of the present invention, the invention provides following technical scheme: a kind of method of explant sterilization in Plant Tissue Breeding, first after adopting liquor potassic permanganate to soak explant material, rinse through running water, then explant material is soaked in thimerosal and ultrasonic echography scheduled time while, finally, with sterile water wash several, complete sterilization.
As preferred embodiment of the present invention, described thimerosal is the mixed solution of clorox and tween disinfectant.
As preferred embodiment of the present invention, it specifically comprises the steps:
Step 1: utilize liquor potassic permanganate immersion explant material 20～40min of 0.025～0.1%, after soaking, material running water rinses explant 30～60min, completes pretreatment;
Step 2: above-mentioned pretreated explant material is immersed in the solution of the clorox that contains 5～10t% and 0.1～0.2% tween disinfectant, and with Vltrasonic device ultrasonic oscillation 4～10min, then use sterile water wash explant material 4～7 times.
As preferred embodiment of the present invention, in described step 1, utilize 0.05% liquor potassic permanganate immersion explant material 30min.
As preferred embodiment of the present invention, in described step 1, in described step 1, the time that running water rinses explant material is 50min.
As preferred embodiment of the present invention, in described step 2, pretreated explant material is immersed in the solution of the clorox that contains 8t% and 0.15% tween disinfectant, and use ultrasonic oscillation 8min.
As preferred embodiment of the present invention, in described step 2, with the number of times of sterile water wash explant material be 7 times.
As preferred embodiment of the present invention, in described step 2, the power of described Vltrasonic device is 150W.
The method that the invention provides 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 above-mentioned purpose of the present invention, feature and advantage can be become apparent more, below the specific embodiment of the present invention is described in detail.
A lot of details are set forth 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 and implement, those skilled in the art can do similar popularization without prejudice to intension of the present invention in the situation that, and therefore the present invention is not subject to the restriction of following public specific embodiment.
The invention provides the method for explant sterilization in a kind of Plant Tissue Breeding, first after adopting liquor potassic permanganate to soak explant material, rinse through running water, then explant material is soaked in thimerosal and ultrasonic echography scheduled time while, finally, with sterile water wash several, 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 of 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 clorox and tween disinfectant.
Embodiment 1
Explant material is cut into segment or fritter or sheet or thread, then carries out disinfection according to the following steps:
Step 1: utilize 0.025% liquor potassic permanganate immersion explant material 40min, after soaking, material running water rinses explant 60min, completes pretreatment;
Step 2: above-mentioned pretreated explant material is immersed in the solution of the clorox that contains 5t% and 0.1% tween disinfectant, and use ultrasonic oscillation 10min, then use sterile water wash explant material 7 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 1: utilize 0.05% liquor potassic permanganate immersion explant material 30min, after soaking, material running water rinses explant 50min, completes pretreatment;
Step 2: above-mentioned pretreated explant material is immersed in the solution of the clorox that contains 8t% and 0.15% tween disinfectant, and use ultrasonic oscillation 8min, then use sterile water wash explant material 7 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 1: utilize 0.1% liquor potassic permanganate immersion explant material 20min, after soaking, material running water rinses explant 30min, completes pretreatment;
Step 2: above-mentioned pretreated explant material is immersed in the solution of the clorox that contains 10t% and 0.1% tween disinfectant, and use ultrasonic oscillation 4min, then use sterile water wash explant material 4 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 1: utilize 0.025% liquor potassic permanganate immersion explant material 20min, after soaking, material running water rinses explant 30min, completes pretreatment;
Step 2: above-mentioned pretreated explant material is immersed in the solution of the clorox that contains 5t% and 0.1% tween disinfectant, and use ultrasonic oscillation 5min, then use sterile water wash explant material 5 times, complete sterilization.
The power of Vltrasonic device described in the present invention is 150W.
Sterilization effect
Select the sterilization method of the embodiment of the present invention 1～4 and traditional sterilizing methods to organize culture experiment to explant, its aseptic rate contrast table is in table 1:
Table 1 sterilization method of the present invention and the traditional method for disinfection killing effect to bacterium and fungi
| Sequence number | Sterilization complexity | Aseptic rate (%) |
| Embodiment 1 | Easily | 97 |
| Embodiment 2 | The easiest | 100 |
| Embodiment 3 | The easiest | 100 |
| Embodiment 4 | Easily | 98 |
| Tradition method for disinfection | Be difficult for | 20.1 |
It should be noted that, kind and the position of the present invention to explant do not limit.
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 unrestricted in order to technical scheme of the present invention to be described, although the present invention is had been described in detail with reference to preferred embodiment, those of ordinary skill in the art is to be understood that, can modify or be equal to replacement technical scheme of the present invention, and not departing from the spirit and scope of technical solution of the present invention, it all should be encompassed in the middle of claim scope of the present invention.
Claims (8)
1. the method for explant sterilization in a Plant Tissue Breeding, it is characterized in that: after first adopting liquor potassic permanganate to soak explant material, rinse through running water, then explant material is soaked in thimerosal and ultrasonic echography scheduled time while, finally, with sterile water wash several, complete sterilization.
2. the method for explant sterilization in Plant Tissue Breeding according to claim 1, is characterized in that: described thimerosal is the mixed solution of clorox and tween disinfectant.
3. the method for explant sterilization in Plant Tissue Breeding according to claim 2, is characterized in that: it specifically comprises the steps:
Step 1: utilize liquor potassic permanganate immersion explant material 20～40min of 0.025～0.1%, after soaking, material running water rinses explant 30～60min, completes pretreatment;
Step 2: above-mentioned pretreated explant material is immersed in the solution of the clorox that contains 5～10t% and 0.1～0.2% tween disinfectant, and with Vltrasonic device ultrasonic oscillation 4～10min, then use sterile water wash explant material 4～7 times.
4. the method for explant sterilization in Plant Tissue Breeding according to claim 3, is characterized in that:
In described step 1, utilize 0.05% liquor potassic permanganate immersion explant material 30min.
5. the method for explant sterilization in Plant Tissue Breeding according to claim 3, is characterized in that: in described step 1, the time that running water rinses explant material is 50min.
6. the method for explant sterilization in Plant Tissue Breeding according to claim 3, it is characterized in that: in described step 2, pretreated explant material is immersed in the solution of the clorox that contains 8t% and 0.15% tween disinfectant, and use ultrasonic oscillation 8min.
7. the method for explant sterilization in Plant Tissue Breeding according to claim 3, is characterized in that: in described step 2, with the number of times of sterile water wash explant material be 7 times.
8. the method for explant sterilization in Plant Tissue Breeding according to claim 3, is characterized in that: in described step 2, the power of described Vltrasonic device is 150W.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108184673A (en) * | 2018-03-08 | 2018-06-22 | 贵州省山地资源研究所 | A kind of tissue culture method of kadsurae coccineae with bud tender stem |
| CN108184672A (en) * | 2018-03-07 | 2018-06-22 | 贵州省山地资源研究所 | A kind of Stauntonia latifolia callus from stem segment method for inducing and cultivating |
| CN108419675A (en) * | 2018-03-13 | 2018-08-21 | 贵州省山地资源研究所 | A kind of tissue culture method of violet passion fruit top tip |
| CN109315289A (en) * | 2018-10-09 | 2019-02-12 | 云南善源生物科技发展有限公司 | Hymsleya amabilis Novel sterilizing mode |
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| CN101743907A (en) * | 2008-12-17 | 2010-06-23 | 宜宾学院 | Rhizoma curculiginis breeding technique |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108184672A (en) * | 2018-03-07 | 2018-06-22 | 贵州省山地资源研究所 | A kind of Stauntonia latifolia callus from stem segment method for inducing and cultivating |
| CN108184672B (en) * | 2018-03-07 | 2021-06-15 | 贵州省山地资源研究所 | Method for inducing and culturing calluses of stem segments of holboellia latifolia |
| CN108184673A (en) * | 2018-03-08 | 2018-06-22 | 贵州省山地资源研究所 | A kind of tissue culture method of kadsurae coccineae with bud tender stem |
| CN108419675A (en) * | 2018-03-13 | 2018-08-21 | 贵州省山地资源研究所 | A kind of tissue culture method of violet passion fruit top tip |
| CN108419675B (en) * | 2018-03-13 | 2021-07-13 | 贵州省山地资源研究所 | Tissue culture seedling raising method for passion fruit top shoots |
| CN109315289A (en) * | 2018-10-09 | 2019-02-12 | 云南善源生物科技发展有限公司 | Hymsleya amabilis Novel sterilizing mode |
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Application publication date: 20141022 |