Comparative analysis of transgenic rice plants obtained by Agrobacterium-mediated transformation and particle bombardment

被引:217
作者
Dai, SH
Zheng, P
Marmey, P
Zhang, SP
Tian, WZ
Chen, SY
Beachy, RN
Fauquet, C
机构
[1] Scripps Res Inst, Res Inst, ILTAB, ORSTOM,TSRI, La Jolla, CA 92037 USA
[2] Chinese Acad Sci, Inst Genet, Beijing 100101, Peoples R China
关键词
agrobacterium; biolistic; rice; transgene analysis;
D O I
10.1023/A:1009687511633
中图分类号
S3 [农学(农艺学)];
学科分类号
0901 ;
摘要
We compared rice transgenic plants obtained by Agrobacterium-mediated and particle bombardment transformation by carrying out molecular analyses of the T-0, T-1 and T-2 transgenic plants. Oryza sativa japonica rice (c.v. Taipei 309) was transformed with a construct (pWNHG) that carried genes coding for neomycin phosphotransferase (nptII), hygromycin phosphotransferase (Hyg(r)), and beta -glucoronidase (GUS). Thirteen and fourteen transgenic lines produced via either method were selected and subjected to molecular analysis. Based on our data, we could draw the following conclusions. Average gene copy numbers of the three transgenes were 1.8 and 2.7 for transgenic plants obtained by Agrobacterium and by particle bombardment, respectively. The percentage of transgenic plants containing intact copies of foreign genes, especially non-selection genes, was higher for Agrobacterium-mediated transformation. GUS gene expression level in transgenic plants obtained from Agrobacterium-mediated transformation was more stable overall the transgenic plant lines obtained by particle bombardment. Most of the transgenic plants obtained from the two transformation systems gave a Mendelian segregation pattern of foreign gens in T-1 and T-2 generations. Co-segregation was observed for liens obtained from particle bombardment, however, that was not always the case for T-1 lines obtained from Agrobacterium-mediated transformation. Fertility of transgenic plants obtained from Agrobacterium-mediated transformation was better. In summary, the Agrobacterium-mediated transformation is a good system to obtain transgenic plants with lower copy number, intact foreign gene and stable gene expression, while particle bombardment is a high efficiency system to produce large number of transgenic plants with a wide range of gene expression.
引用
收藏
页码:25 / 33
页数:9
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