Agrobacterium-mediated sorghum transformation

被引:206
作者
Zhao, ZY [1 ]
Cai, TS [1 ]
Tagliani, L [1 ]
Miller, M [1 ]
Wang, N [1 ]
Pang, H [1 ]
Rudert, M [1 ]
Schroeder, S [1 ]
Hondred, D [1 ]
Seltzer, J [1 ]
Pierce, D [1 ]
机构
[1] Pioneer Hi Bred Int Inc, Trait & Technol Dev, Johnston, IA 50131 USA
关键词
Agrobacterium tumefaciens; sorghum; transformation; transgenic plants;
D O I
10.1023/A:1026507517182
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Agrobacterium tumefaciens was used to genetically transform sorghum. Immature embryos of a public (P898012) and a commercial line (PHI391) of sorghum were used as the target explants. The Agrobacterium strain used was LBA4404 carrying a 'Super-binary' vector with a bar gene as a selectable marker for herbicide resistance in the plant cells. A series of parameter tests was used to establish a baseline for conditions to be used in stable transformation experiments. A number of different transformation conditions were tested and a total of 131 stably transformed events were produced from 6175 embryos in these two sorghum lines. Statistical analysis showed that the source of the embryos had a very significant impact on transformation efficiency, with field-grown embryos producing a higher transformation frequency than greenhouse-grown embryos. Southern blot analysis of DNA from leaf tissues of T-0 plants confirmed the integration of the T-DNA into the sorghum genome. Mendelian segregation in the T-1 generation was confirmed by herbicide resistance screening. This is the first report of successful use of Agrobacterium for production of stably transformed sorghum plants. The Agrobacterium method we used yields a higher frequency of stable transformation that other methods reported previously.
引用
收藏
页码:789 / 798
页数:10
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