CN1379970A - Soybean transgene plant cultur method - Google Patents
Soybean transgene plant cultur method Download PDFInfo
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- CN1379970A CN1379970A CN 02111705 CN02111705A CN1379970A CN 1379970 A CN1379970 A CN 1379970A CN 02111705 CN02111705 CN 02111705 CN 02111705 A CN02111705 A CN 02111705A CN 1379970 A CN1379970 A CN 1379970A
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- msb
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Abstract
The soybean transgenic plant culture method includes the following steps: placing advertitions bud into solid MSB bud elongating culture medium containing 50 mg/L resistomycin, at the same time adding liquid bud elongating culture medium, and placing the transformed bud into the solid MSB rooting culture medium containing 30 mg/L resistomycin, at the same time adding liquid rooting culture medium, and using liquid transformation seedling culture liquor to make culture so as to obtain the transgenic soybean seed system. Said invention adopts the combination mode of solid culture medium and liquid culture medium, and adds a certain quantity of resistomycin so as to acelerate soybean plant to root and sprout, and can inhibit growth of chimera, and can make plant quickly grow and pod more.
Description
Technical field:
The present invention relates to a kind of soybean transgene plant cultur method, is a kind of technology of biological culture, belongs to biotechnology and modern agricultural technology field.
Background technology:
By transgenic technology the external source genes of interest is imported in the acceptor soybean explant, tissue culture and plant regeneration through transformant, cultivate the new varieties with foreign gene characteristic, the cultivation of soybean indefinite bud therebetween, the growth of taking root and transforming seedling that transforms bud are the key links in the culture technology.Prior art all is to adopt MSB (MS (Murashige and Skoog) inorganic constituents+B5 (Gamborg) organic principle) solid culture medium (" modern plants Physiology Experiment guide " Tang Zhangcheng chief editor, Science Press in cultivation, 1999), because solid culture medium and the plant tissue contact area is few, gas exchange is not smooth, nutriment concentration is variant and cause the imbalance of growth easily.
Summary of the invention:
The objective of the invention is to overcome the deficiency of existing solid culture medium, a kind of soybean transgene plant cultur method is provided, solve to transform the low and poky difficult point of tissue cultivating seedling of screening accuracy rate of seedling, reach chimera few, take root soon, robust growth improves the bear pods effect of rate of seedling.
For realizing such purpose, in the technical scheme of the present invention, adopt solid culture medium and the liquid nutrient medium best cultivation that mutually combines, adopt bud elongation medium and root media respectively in different vegetative period, and in medium, add a certain amount of kanamycin sulfate, make soybean plant strain take root, germinate, growth rapidly.
Concrete grammar of the present invention comprises the steps:
1, cuts the sprouting that comes from the slough, the soybean that the forms indefinite bud of growing thickly is put on the solid MSB bud elongation medium, and add the high liquid MSB bud elongation medium of 0.1～0.3cm simultaneously, solid and liquid bud elongation medium all contain the 50mg/L kanamycin sulfate, and concrete composition is: MSB+0.2mg/LBA+0.2mg/LIBA+0.3mg/LGA
3+ Km50mg/L, illumination 16 hours every days is in 26 ℃ of cultivations 15～20 days, to transforming bud 3～4cm.
2, the conversion bud of the about 3～4cm of height is downcut, put on the solid MSB root media, and add the high liquid MSB root media of 0.5～0.7cm simultaneously, solid and liquid root media all contain the 30mg/L kanamycin sulfate, concrete composition is 1/2MSB+1.5mg/LIBA+Km30mg/L, cultivated for 2～3 weeks, root induction obtains transforming seedling.
3, the conversion seedling about will long 5cm adds 0.9cm liquid MSB medium, and the composition of liquid MSB medium is MSB+1.5mg/LIBA+Km25mg/L, the macroelement of liquid MSB medium: NH
4NO
3, KNO
3, KH
2PO
4, MgSO
47H
2O, CaCl
22H
2The equal decrement 70%～80% of O after the conversion seedling is bloomed and born pods, can obtain genetically engineered soybean kind system.
The present invention has substantive distinguishing features and marked improvement, compares with common solid culture medium, and solid and liquid culture mutually combine and more can suppress chimeric growth, helps taking root, and forms more hair; Transfer-gen plant solid and liquid culture binding energy make plant strain growth fast, bear pods more.
Embodiment:
Below in conjunction with specific embodiment technical scheme of the present invention is further described.
Embodiment 1
The test kind is for closing rich 35
Transforming bud elongation medium composition is:
MSB+0.2mg/LBA+0.2mg/LIBA+0.3mg/LGA
3+Km50mg/L，
Adopt the high liquid nutrient medium of solid+0.3cm, cultivated 15 days, transform bud and be stretched to 3～4cm;
Transforming bud root induction medium component is:
1/2MSB+1.5mg/LIBA+Km25mg/L，
Adopt the high liquid nutrient medium of solid+0.5cm, cultivated 14 days, obtain the soybean transgene resistant plant;
The transfer-gen plant medium component is:
MSB+1.5mg/LIBA+Km25mg/L，
Adopt the high liquid nutrient medium of solid+0.8cm, NH
4NO
3, KNO
3, KH
2PO
4, MgSO
47H
2O, CaCl
22H
2The equal decrement 80% of O is bloomed and is born pods, and obtains genetically engineered soybean kind system.
Embodiment 2
The test kind is for closing rich 35
Transforming bud elongation medium composition is:
MSB+0.2mg/LBA+0.2mg/LIBA+0.3mg/LGA
3+Km50mg/L，
Adopt the high liquid nutrient medium of solid+0.2cm, cultivated 18 days, transform bud and be stretched to 3～4cm;
Transforming bud root induction medium component is:
1/2MSB+1.5mg/LIBA+Km25mg/L，
Adopt the high liquid nutrient medium of solid+0.6cm, cultivated 18 days, obtain the soybean transgene resistant plant;
The transfer-gen plant medium component is:
MSB+1.5mg/LIBA+Km25mg/L，
Adopt the high liquid nutrient medium of solid+0.9cm, NH
4NO
3, KNO
3, KH
2PO
4, MgSO
47H
2O, CaCl
22H
2The equal decrement 70% of O is bloomed and is born pods, and obtains genetically engineered soybean kind system.
Embodiment 3
The test kind is for closing rich 35
Transforming bud elongation medium composition is:
MSB+0.2mg/LBA+0.2mg/LIBA+0.3mg/LGA
3+Km50mg/L，
Adopt the high liquid nutrient medium of solid+1.0cm, cultivated 20 days, transform bud and be stretched to 3～4cm;
Transforming bud root induction medium component is:
1/2MSB+1.5mg/LIBA+Km25mg/L，
Adopt the high liquid nutrient medium of solid+0.7cm, cultivated 21 days, obtain the soybean transgene resistant plant;
The transfer-gen plant medium:
MSB+1.5mg/LIBA+Km25mg/L，
Adopt the high liquid nutrient medium of solid+0.1cm, NH
4NO
3, KNO
3, KH
2PO
4, MgSO
47H
2O, CaCl
22H
2The equal decrement 75% of O is bloomed and is born pods, and obtains genetically engineered soybean kind system.
With the method plant strain growth is speeded up, healthy and strong more, biological yield increases.
Claims (1)
1, a kind of soybean transgene plant cultur method is characterized in that specifically comprising the steps:
1. the indefinite bud that forms is put on the solid MSB bud elongation medium, added the high liquid MSB bud elongation medium of 0.1～0.3cm simultaneously, MSB bud elongation medium composition is MSB+0.2mg/LBA+0.2mg/LIBA+0.3mg/LGA
3+ Km50mg/L, illumination 16 hours every days is in 26 ℃ of cultivations 15～20 days, to transforming bud 3～4cm;
2. downcut transforming bud, broadcast on the solid MSB root media, add the high liquid MSB root media of 0.5～0.7cm simultaneously, the composition of root media is 1/2MSB+1.5mg/LIBA+Km25mg/L, cultivates for 2～3 weeks, and root induction must transform seedling;
3. will transform seedling and add 0.9cm liquid MSB medium, the composition of liquid MSB medium is MSB+1.5mg/LIBA+Km25mg/L, wherein macroelement: NH
4NO
3, KNO
3, KH
2PO
4, MgSO
47H
2O, CaCl
22H
2The equal decrement 70%～80% of O obtains genetically engineered soybean kind system.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021117055A CN1166284C (en) | 2002-05-16 | 2002-05-16 | Soybean transgene plant cultur method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021117055A CN1166284C (en) | 2002-05-16 | 2002-05-16 | Soybean transgene plant cultur method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1379970A true CN1379970A (en) | 2002-11-20 |
| CN1166284C CN1166284C (en) | 2004-09-15 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021117055A Expired - Fee Related CN1166284C (en) | 2002-05-16 | 2002-05-16 | Soybean transgene plant cultur method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101405394B (en) * | 2006-03-17 | 2012-10-10 | 巴斯福植物科学有限公司 | D-amino acid selection for soybean |
| CN106978435A (en) * | 2016-01-18 | 2017-07-25 | 中国科学院上海生命科学研究院 | A kind of method for the transgenosis frequency for improving soybean |
-
2002
- 2002-05-16 CN CNB021117055A patent/CN1166284C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101405394B (en) * | 2006-03-17 | 2012-10-10 | 巴斯福植物科学有限公司 | D-amino acid selection for soybean |
| CN106978435A (en) * | 2016-01-18 | 2017-07-25 | 中国科学院上海生命科学研究院 | A kind of method for the transgenosis frequency for improving soybean |
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| Publication number | Publication date |
|---|---|
| CN1166284C (en) | 2004-09-15 |
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