Genetic transformation of Coffea canephora by vacuum infiltration

被引:25
|
作者
Canche-Moo, RLR
Ku-Gonzalez, A
Burgeff, C
Loyola-Vargas, VM
Rodríguez-Zapata, LC
Castaño, E
机构
[1] Ctr Invest Cient Yucatan, Unidad Bioquim & Biol Mol Plantas, Merida 97200, Yucatan, Mexico
[2] Ctr Invest Cient Yucatan, Unidad Biotecnol, Merida 97200, Yucatan, Mexico
关键词
Agrobacterium transformation; coffee; somatic embryogenesis; vacuum infiltration;
D O I
10.1007/s11240-005-9036-4
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Agrobacterium-mediated plant transformation protocol was evaluated as a fast method to obtain genetically modified Coffea canephora plantlets. Leaf explants were used as source material for Agrobacterium tumefaciens-mediated transformation involving a vacuum infiltration protocol, followed by a step of somatic embryogenesis induction and a final selection of the transformed plants. A. tumefaciens strain C58CI containing the binary vector pER10W-35SRed was used. PCR amplification of DsRFP gene and visual detection of the red fluorescent protein demonstrated 33% transformed embryos. The protocol presented here produces reliable transgenic coffee embryos in two months.
引用
收藏
页码:373 / 377
页数:5
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