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2011•Gansu Agricultural Science and TechnologyRequires access

Optimization of the High Efficient Genetic Transformation for Tobacco NR Mediated by Agrobacterium

Yuliang Chen

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Abstract

A rapid and high efficient transformation system has been established for Agrobacterium-mediated transformation of tobacco by optimizing the factors of transformation.The tobacco leaves were cut into 1.0 cm×1.0 cm discs and then were submerged in Agrobacterium suspensions(OD600=0.5) for 5～10 min,then were transferred to MS differentiation medium supplemented with 2.25 mg/L 6-BA and 0.10 mg/LNAA.The buds resistant to kanamycin were obtained in 21～28 days under 100 mg/L kanamycin selection.The positive rates of putative transgenic plants could reach 81.2% by using rooting selection of kanamycin and PCR identification.

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What this paper is about

A rapid and high efficient transformation system has been established for Agrobacterium-mediated transformation of tobacco by optimizing the factors of transformation.The tobacco leaves were cut into 1.0 cm×1.0 cm discs and then were submerged in Agrobacterium suspensions(OD600=0.5) for 5～10 min,then were transferred to MS differentiation medium supplemented with 2.25 mg/L 6-BA and 0.10 mg/LNAA.The buds resistant to kanamycin were obtained in 21～28 days under 100 mg/L kanamycin selection.The positive rates of putative transgenic plants could reach 81.2% by using rooting selection of kanamycin and PCR identification.

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Available abstract

A rapid and high efficient transformation system has been established for Agrobacterium-mediated transformation of tobacco by optimizing the factors of transformation.The tobacco leaves were cut into 1.0 cm×1.0 cm discs and then were submerged in Agrobacterium suspensions(OD600=0.5) for 5～10 min,then were transferred to MS differentiation medium supplemented with 2.25 mg/L 6-BA and 0.10 mg/LNAA.The buds resistant to kanamycin were obtained in 21～28 days under 100 mg/L kanamycin selection.The positive rates of putative transgenic plants could reach 81.2% by using rooting selection of kanamycin and PCR identification.

Key concepts: Kanamycin, Agrobacterium, Transformation (genetics), Genetically modified crops, Biology, Transgene, Selection (genetic algorithm), Biotechnology

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