| Autor: |
Zhao, Zuo-yu, Gu, Weining, Cai, Tishu, Tagliani, Laura, Hondred, David, Bond, Diane, Schroeder, Sheryl, Rudert, Marjorie, Pierce, Dottie |
| Zdroj: |
Molecular Breeding; December 2002, Vol. 8 Issue: 4 p323-333, 11p |
| Abstrakt: |
A high throughput genetic transformation system in maize has been developed with Agrobacterium tumefaciensmediated T-DNA delivery. With optimized conditions, stable callus transformation frequencies for Hi-II immature embryos averaged approximately 40%, with results in some experiments as high as 50%. The optimized conditions include N6 medium system for Agrobacteriuminoculation, co-cultivation, resting and selection steps; no AgNo3in the infection medium and adding AgNo3in co-cultivation, resting and selection medium; Agrobacteriumconcentration at 0.5×109c.f.u. ml−1for bacterium inoculation; 100 mg l−1carbenicillin used in the medium to eliminate Agrobacteriumafter inoculation; and 3 days for co-cultivation and 4 days for resting. A combination of all of these conditions resulted in establishing a high throughput transformation system. Over 500 T0plants were regenerated and these plants were assayed by transgene expression and some of them were also analyzed by Southern hybridization. T1plants were analyzed and transmission of transgenes to the T1generation was verified. This represents a highly reproducible and reliable system for genetic transformation of maize Hi-II. |
| Databáze: |
Supplemental Index |
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