CN1406460A - Method for disinfecting transgenic soybean seeds - Google Patents
Method for disinfecting transgenic soybean seeds Download PDFInfo
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- CN1406460A CN1406460A CN 02150780 CN02150780A CN1406460A CN 1406460 A CN1406460 A CN 1406460A CN 02150780 CN02150780 CN 02150780 CN 02150780 A CN02150780 A CN 02150780A CN 1406460 A CN1406460 A CN 1406460A
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- mercury chloride
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Abstract
A method for disinfecting the transgenic seeds of soybean to kill different microbes features that the transgenic seeds are disinfected in the disinfecting liquid at 2-6 deg.C for 50-70 min. Its advantages are thorough disinfection, high germination percentaeg and robust seedlings.
Description
Technical field
The present invention relates to a kind of seed disinfection processing method, particularly a kind of soybean transgene seed disinfection processing method belongs to biotechnology and modern agricultural technology field.
Background technology
Along with development of biology, genetically modified crops gradually move towards market, soybean is one of them important crop.Seed disinfection is first link of work in soyabean tissue cultivates, and pathogen contamination is the big obstacle that sterile tissue is cultivated.Find that by literature search Gansu science tech publishing house published in 1996, " practical plant tissue culture technique study course " P39 page or leaf of chief editors such as Cao Ziyi: conventional method is generally used 0.1%HgCl
2Solution soaking 15 minutes or saturated Eusol soaked 25 minutes.But usually sterilization is not thorough at work, and pollution happens occasionally, and causes the accuracy of the waste and the influence experiment of test material, if the time soaks long then seed sprouting is caused infringement, percentage of seedgermination is descended, and germination vigor weakens.
Summary of the invention and embodiment
The objective of the invention is to overcome the deficiency and the defective of existing sterilization method, a kind of soybean transgene seed disinfection processing method is provided, make the soya seeds sterilization thoroughly, pollution-free, do not influence seed germination again, the seedling robust growth, it is many to reach soybean group training indefinite bud, the effect of fast growth.The present invention is achieved through the following technical solutions: the present invention mainly adopts the mode of control temperature, temperature is controlled at 2 ℃～6 ℃, after adding thimerosal, and prolong disinfecting time to 50～70 minute, thereby kill various microorganisms, can guarantee that the soya seeds pollution rate drops to 0.1%-0.7%, and do not influence the normal sprouting of soya seeds, germination rate remains on 97%-99%, and germination vigor is strong.
Below the invention will be further described, method step is as follows:
(1) soya seeds, 75% alcohol, 0.1% mercury chloride or saturated Eusol remain on 2 ℃～6 ℃.
(2) select kind of a skin and do not have breakage, no scab seed cleans up through running water, blocked down in alcohol-pickled 30 seconds with 2 ℃～6 ℃ 75% again and do.
(3) add 2 ℃～6 ℃ 0.1% mercury chloride or saturated Eusol and soak, and remained in the scope of 2 ℃～6 ℃ of temperature 50～70 minutes.
(4) under aseptic technique, add to be connected on the MS medium after 2 ℃～6 ℃ sterile water 500ml flushing 3～8 times and sprout, through cultivating the seedling that obtained sprouting in 5～7 days.
The present invention has substantive distinguishing features and marked improvement, the invention solves the pollution that seed does not thoroughly cause owing to sterilization, and owing to disinfecting time length influences the soya seeds germination rate, weaken the difficult point of germination vigor, the soya seeds pollution rate is descended, and keep high germination rate and strong germination vigour, and select 0.1%HgCl below the normal temperature for use
2Solution soaking 15 minutes or saturated Eusol soak to be compared in 25 minutes, and seed disinfection is thorough, and germination rate rises germination vigor enhancing, robust growth.
Provide following three embodiment in conjunction with content of the present invention:
Embodiment 1-experiment kind is for closing rich 35
Soya seeds, 75% alcohol, 0.1% mercury chloride or saturated Eusol keep 2 ℃, choose kind of a skin and do not have breakage, no scab seed cleans up through running water, block down dried in alcohol-pickled 30 seconds with 2 ℃ 70% again; Add 2 ℃ of 1% mercury chloride or saturated Eusol and soak, and remained in the scope of 2 ℃ of temperature 50 minutes; Under aseptic technique, add to be connected on the MS medium after 2 ℃ of sterile water 500ml wash 3 times and sprouted 5 days.The experiment kind is closed rich 35 pollution rates 0.7%, germination rate 97%, and germination vigor is strong.
Embodiment 2-experiment kind is for closing rich 25
Soya seeds, 75% alcohol, 0.1% mercury chloride or saturated Eusol keep 4 ℃, plant that skin does not have breakage, no scab seed cleans up through running water, block down dried in alcohol-pickled 30 seconds with 4 ℃ 70% again; Add 4 ℃ of 1% mercury chloride or saturated Eusol and soak, and remained in the scope of 4 ℃ of temperature 60 minutes; Under aseptic technique, add to be connected on the MS medium after 4 ℃ of sterile water 500ml wash 6 times and sprouted 6 days.The experiment kind is closed rich 25 pollution rates 0.5%.Germination rate 99%, germination vigor is strong.
Embodiment 3-experiment kind is for closing rich 39
Soya seeds, 75% alcohol, 0.1% mercury chloride or saturated Eusol keep 6 ℃, plant that skin does not have breakage, no scab seed cleans up through running water, block down dried in alcohol-pickled 30 seconds with 6 ℃ 70% again; Add 6 ℃ of 1% mercury chloride or saturated Eusol and soak, and remained in the scope of 6 ℃ of temperature 70 minutes; Under aseptic technique, add to be connected on the MS medium after 6 ℃ of sterile water 500ml wash 8 times and sprouted 7 days.The experiment kind is closed rich 39 pollution rates 0.1%, germination rate 98%, and germination vigor is strong.
Claims (2)
1, a kind of soybean transgene seed disinfection processing method is characterized in that: adopt the mode of control temperature, temperature is controlled at 2 ℃～6 ℃, after adding thimerosal, and prolong disinfecting time to 50～70 minute, thereby kill various microorganisms.
2, this soybean transgene seed disinfection processing method according to claim 1 is characterized in that method step is as follows:
(1) soya seeds, 75% alcohol, 0.1% mercury chloride or saturated Eusol keep 2 ℃～6 ℃;
(2) selecting kind of a skin does not have breakage, no scab seed, cleans for several times through tap water, use 2 ℃～6 ℃ aseptic water washings for several times again after, with 2 ℃～6 ℃ 70% alcohol-pickled 30 seconds, block down dried;
(3) add 2 ℃～6 ℃ 0.1% mercury chloride or saturated Eusol and soak, and remained in the scope of 2 ℃～6 ℃ of temperature 50～70 minutes;
(4) under aseptic technique, add to be connected to after 2 ℃～6 ℃ sterile water 500ml flushing 3～8 times and sprouted on the MS medium 5～7 days, the seedling that obtains sprouting through cultivation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021507805A CN1172569C (en) | 2002-11-28 | 2002-11-28 | Method for disinfecting transgenic soybean seeds |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021507805A CN1172569C (en) | 2002-11-28 | 2002-11-28 | Method for disinfecting transgenic soybean seeds |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1406460A true CN1406460A (en) | 2003-04-02 |
| CN1172569C CN1172569C (en) | 2004-10-27 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021507805A Expired - Fee Related CN1172569C (en) | 2002-11-28 | 2002-11-28 | Method for disinfecting transgenic soybean seeds |
Country Status (1)
| Country | Link |
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| CN (1) | CN1172569C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103891454A (en) * | 2014-03-24 | 2014-07-02 | 安徽华夏农业科技股份有限公司 | Method for increasing germination rate of soybean seeds |
| CN110073976A (en) * | 2019-05-07 | 2019-08-02 | 山西省农业科学院棉花研究所 | A kind of sterilization method of tissue culture soya seeds |
-
2002
- 2002-11-28 CN CNB021507805A patent/CN1172569C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103891454A (en) * | 2014-03-24 | 2014-07-02 | 安徽华夏农业科技股份有限公司 | Method for increasing germination rate of soybean seeds |
| CN103891454B (en) * | 2014-03-24 | 2015-06-17 | 安徽华夏农业科技股份有限公司 | Method for increasing germination rate of soybean seeds |
| CN110073976A (en) * | 2019-05-07 | 2019-08-02 | 山西省农业科学院棉花研究所 | A kind of sterilization method of tissue culture soya seeds |
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| Publication number | Publication date |
|---|---|
| CN1172569C (en) | 2004-10-27 |
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