Development of transgenic rice pure lines by particle bombardment-mediated transformation and genetic analysis-based selection
文献类型: 外文期刊
作者: Tang, K 1 ; Wu, A 2 ; Yao, J 2 ; Qi, H 2 ; Lu, X 2 ;
作者机构: 1.Fudan Univ, Inst Genet, Sch Life Sci, Shanghai 200433, Peoples R China
2.Fudan Univ, Inst Genet, Sch Life Sci, Shanghai 200433, Peoples R China; Shanghai Jiao Tong Univ, Sch Agr, Shanghai 201101, Peoples R China; Hubei Acad Agr Sci, Hybrid Rice Res Ctr, Wuhan 430064, Hubei, Peoples R China
关键词: Snowdrop lectin. Plants. Expression. Resistance. Acceleration. Dna.;Cotransformation. Gna.
期刊名称:ACTA BIOTECHNOLOGICA ( 影响因子:1.257; )
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收录情况: SCI
摘要: Mature seed-derived callus from an elite Chinese japonica rice cv. Eyl 105 was transformed with a plasmid containing the selectable marker hygromycin phosphotransferase (hpt) and the reporter #beta#-glucuronidase (gusA) genes via particle bombardment. After two rounds of selection on hygromycin (30 mg/l)-containing medium, resistant callus was transferred to hygromycin (30 mg/l)-containing regeneration medium for plant regeneration. Twenty-three independent transgenic rice plants were regenerated from 127 bombarded callus with a transformation frequency of 18.1%. All the transgenic plants contained both gusA and hpt genes, revealed by PCR/Southern blot analysis. GUS assay revealed 18 out of 23 plants (78.3%) proliferated on hygromycin-containing medium had GUS expression at various levels. Genetic analysis confirmed Mendelian segregation of transgenes in progeny. From R2 generations with their R1 parent plants showing 3:1 Mendelian segregation, we identified three independent homozygous transgenic rice lines. The homozygous lines were phenotypically normal and fertile compared to the control plants. We demonstrate that homozygous transgenic rice lines can be obtained via particle bombardment-mediated transformation and through genetic analysis-based selection.
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