Abstract |
In the present study, an efficient Agrobacterium-mediated gene transformation system was developed for soybean [Glycine max (L.) Merrill] based on the examinations of several factors affecting plant transformation efficiency. Increased transformation efficiencies were obtained when the soybean cotyledonary node were inoculated with the Agrobacterium inoculum added with 0.02% (v/v) surfactant (Silwet L-77). The applications of Silwet L-77 (0.02%) during infection and L-cysteine (600 mg l(-1)) during co-cultivation resulted in more significantly improved transformation efficiency than each of the two factors alone. The optimized temperature for infected explant co-cultivation was 22 degrees C. Regenerated transgenic shoots were selected and produced more efficiently with the modified selection scheme (initiation on shoot induction medium without hygromycin for 7 days, with 3 mg l(-1) hygromycin for 10 days, 5 mg l(-1) hygromycin for another 10 days, and elongation on shoot elongation medium with 8 mg l(-1) hygromycin). Using the optimized system, we obtained 145 morphologically normal and fertile independent transgenic plants in five important Chinese soybean varieties. The transformation efficacies ranged from 3.8 to 11.7%. Stable integration, expression and inheritance of the transgenes were confirmed by molecular and genetic analysis. T(1) plants were analyzed and transmission of transgenes to the T(1 )generation in a Mendelian fashion was verified. This optimized transformation system should be employed for efficient Agrobacterium-mediated soybean gene transformation.
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Authors | Sheng-Jun Liu, Zhi-Ming Wei, Jian-Qiu Huang |
Journal | Plant cell reports
(Plant Cell Rep)
Vol. 27
Issue 3
Pg. 489-98
(Mar 2008)
ISSN: 0721-7714 [Print] Germany |
PMID | 18004571
(Publication Type: Journal Article)
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Chemical References |
- Cinnamates
- Hygromycin B
- hygromycin A
- Cysteine
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Topics |
- Blotting, Southern
- Cinnamates
(pharmacology)
- Cysteine
(pharmacology)
- Hygromycin B
(analogs & derivatives, pharmacology)
- Plants, Genetically Modified
- Polymerase Chain Reaction
- Rhizobium
(genetics)
- Soybeans
(drug effects, genetics, growth & development)
- Temperature
- Transformation, Genetic
(drug effects, genetics)
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