Nghiên cứu chuyển gen vào dòng ca cao thương mại TD28 ở Việt Nam thông qua vi khuẩn Agrobacterium tumefaciens

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Tác giả: Hồng Hạnh Hà, Thị Thu Hiền Lê, Văn Hải Nông

Ngôn ngữ: vie

Ký hiệu phân loại: 571.538 *Tissue culture

Thông tin xuất bản: Công nghệ sinh học, 2014

Mô tả vật lý: 255-260

Bộ sưu tập: Metadata

ID: 503894

Cacao is one of impotant industry trees. The development of transgenic cacao plants for genetic improvement has been carried out. In this study, an Agrobacterium-mediated genetic transformation protocol for the commercial TD8 cacao clone has been established. Genes GUSPlus or GUS under the control of promoter 35S or FMV34S were transformed into cotyledon ftagments isolated from mature primary somatic embryos using A. tumefaciens strain EHA105. Infected explants were cocultivated for 2 days on secondary callus growth (SCG) medium containing 2 mgl/12.4-D and 0.05 mg/l BAP, 0.1 mM acetosyringone at 25°C in the dark. The explants formed embryogenic callus on SCG medium supplemented with 50 mg/l kanamycin for NPTII selection and 200 mg/l moxalactarn for eliminating A. tumefaciens. Secondary somatic embryos were induced by transfering of the selected embryogenic callus onto embryo development (ED) medium containing 30 g/l sucrose, 1 g/l glucose, 50 mg/l kanamycin, 200 mg/l moxalactam. Secondary somatic embryos were matured on the embryo development in light (EDL) medium containing 0.3 g/l KNO3, 1 m/l amino acid, 10 g/l sucrose, 20 g/l glucose, 50 mg/l kanamycin, 200 mg/l moxalactarn. Shoot - producing embryos with leaves were transferred to the root development (RD) medium containing 0.3 g/l KNO3, 5 g/l sucrose and 10 g/l glucose, 50 mg/l kanamycin, 200 mg/l moxalactarn. The analysis of GUS expression and PCR of NPTII gene demonstrated the presence of interested genes in transgenic cacao plants.
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