CN105454045A - Disinfection method of grape explant in tissue culture - Google Patents
Disinfection method of grape explant in tissue culture Download PDFInfo
- Publication number
- CN105454045A CN105454045A CN201510878453.9A CN201510878453A CN105454045A CN 105454045 A CN105454045 A CN 105454045A CN 201510878453 A CN201510878453 A CN 201510878453A CN 105454045 A CN105454045 A CN 105454045A
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- CN
- China
- Prior art keywords
- stem section
- grape
- grape stem
- explant
- washed
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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
Abstract
The invention discloses a disinfection method of a grape explant in tissue culture. The disinfection method comprises the following steps: a grape stem section is washed with sterile water for 3-5 times and then washed with 75% alcohol for 30-50 seconds, and a grape stem section after alcohol washing is obtained; the grape stem section is washed with the sterile water for 3-5 times and soaked in 800-1,000 times of the 84 disinfectant for 2-4 minutes, and the grape stem section after 84 disinfectant soaking is obtained; the grape stem section is washed with the sterile water for 3-5 times and soaked in 0.1% mercuric chloride for 5-8 minutes, and the grape stem section after mercuric chloride soaking is obtained; the grape stem section is washed with the sterile water for 3-5 times, and disinfection is finished for inoculation. The disinfection method has the following benefits: after the disinfection method of the grape explant in tissue culture is adopted for disinfection, the explant is inoculated to a medium for culture, the mould generation rate after rooting and leafing can be reduced by more than 60%, and accordingly, the method can effectively reduce mould contamination of the explant after inoculation.
Description
Technical field
The present invention relates to plant tissue explant disinfection method field, be specifically related to the sterilization method that grapes tissue cultivates explant.
Background technology
The a large amount of acquisition time of grape explant is annual July, if rainwater is more, disease can be very heavy, even if there is no mould symptom on the grape leave of land for growing field crops, also recessiveness can carry disease germs in plant body, treat that explant is inoculated into medium 10-15 days, explant has been taken root Zhan Yehou, after plant cell division, in plant strain growth process, inherent mould contamination produces.With the very difficult sterilization of conventional sterilization method at present thorough (general explant inoculated and cultured, mould contamination occurs within 3 days after inoculation).
Summary of the invention
Object of the present invention is exactly for above-mentioned defect of the prior art, provides the sterilization method that grapes tissue cultivates explant.
To achieve these goals, technical scheme provided by the invention is: grapes tissue cultivates the sterilization method of explant, comprises the following steps:
1) by grape stem section with after aseptic water washing 3-5 time, then with 75% alcohol rinse 30-50 second, obtain grape stem section after alcohol rinse;
2) grape stem section after alcohol rinse step 1) obtained, after aseptic water washing 3-5 time, then with 800-1000 doubly 84 medicining liquid dipping 2-4 minute, obtains grape stem section after 84 medicining liquid dippings;
3) by step 2) grape stem section after 84 medicining liquid dippings that obtain, after aseptic water washing 3-5 time, then soak 5-8 minute with 0.1% mercuric chloride, obtain mercuric chloride soak after grape stem section;
4) after mercuric chloride step 3) obtained soaks, grape stem section aseptic water washing 3-5 time, namely sterilizes complete, to be seeded.
Beneficial effect of the present invention is: the sterilization method adopting grapes tissue provided by the invention to cultivate explant carries out disinfection, explant is inoculated into medium and cultivates, the mould after opening up leaf that takes root occurs to reduce by more than 60%, illustrates that the method can effectively reduce the rear mould contamination of explant inoculation.
Embodiment
embodiment 1:
Grapes tissue cultivates the sterilization method of explant, comprises the following steps:
1) by grape stem section with after aseptic water washing 3-5 time, then with 75% alcohol rinse 30-50 second, obtain grape stem section after alcohol rinse;
2) grape stem section after alcohol rinse step 1) obtained, after aseptic water washing 3-5 time, then with 800-1000 doubly 84 medicining liquid dipping 2-4 minute, obtains grape stem section after 84 medicining liquid dippings;
3) by step 2) grape stem section after 84 medicining liquid dippings that obtain, after aseptic water washing 3-5 time, then soak 5-8 minute with 0.1% mercuric chloride, obtain mercuric chloride soak after grape stem section;
4) after mercuric chloride step 3) obtained soaks, grape stem section aseptic water washing 3-5 time, namely sterilizes complete, to be seeded.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (1)
1. grapes tissue cultivates the sterilization method of explant, it is characterized in that, comprises the following steps:
1) by grape stem section with after aseptic water washing 3-5 time, then with 75% alcohol rinse 30-50 second, obtain grape stem section after alcohol rinse;
2) grape stem section after alcohol rinse step 1) obtained, after aseptic water washing 3-5 time, then with 800-1000 doubly 84 medicining liquid dipping 2-4 minute, obtains grape stem section after 84 medicining liquid dippings;
3) by step 2) grape stem section after 84 medicining liquid dippings that obtain, after aseptic water washing 3-5 time, then soak 5-8 minute with 0.1% mercuric chloride, obtain mercuric chloride soak after grape stem section;
4) after mercuric chloride step 3) obtained soaks, grape stem section aseptic water washing 3-5 time, namely sterilizes complete, to be seeded.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510878453.9A CN105454045A (en) | 2015-12-04 | 2015-12-04 | Disinfection method of grape explant in tissue culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510878453.9A CN105454045A (en) | 2015-12-04 | 2015-12-04 | Disinfection method of grape explant in tissue culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105454045A true CN105454045A (en) | 2016-04-06 |
Family
ID=55592616
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510878453.9A Pending CN105454045A (en) | 2015-12-04 | 2015-12-04 | Disinfection method of grape explant in tissue culture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105454045A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102742507A (en) * | 2012-08-03 | 2012-10-24 | 贵州大学 | In-vitro regeneration culture method for downy grape |
| CN103190345A (en) * | 2013-04-14 | 2013-07-10 | 巴中市光雾山植物研究所 | One-step tissue culture and rapid propagation method of grapes |
| CN104542298A (en) * | 2015-01-26 | 2015-04-29 | 武汉市林业果树科学研究所 | Grape tissue regeneration culture method |
-
2015
- 2015-12-04 CN CN201510878453.9A patent/CN105454045A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102742507A (en) * | 2012-08-03 | 2012-10-24 | 贵州大学 | In-vitro regeneration culture method for downy grape |
| CN103190345A (en) * | 2013-04-14 | 2013-07-10 | 巴中市光雾山植物研究所 | One-step tissue culture and rapid propagation method of grapes |
| CN104542298A (en) * | 2015-01-26 | 2015-04-29 | 武汉市林业果树科学研究所 | Grape tissue regeneration culture method |
Non-Patent Citations (1)
| Title |
|---|
| 张爱华等: "葡萄砧木—抗砧3号组织培养快繁技术初探", 《新疆农垦科技》 * |
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| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160406 |
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| RJ01 | Rejection of invention patent application after publication |