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| Optimization of Transformation System for Malus baccata |
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Abstract In order to optimize the genetic transformation system for Malus baccata Borkh., DNA recombination technology was used for inserting green fluorescence protein gene (GFP) to pBI121, and then a new recombinant plasmid pBI121-eGFP was constructed. The plasmid was transferred to M. baccata Borkh. after introduced into Agrobacterium tumefaciens EHA105. Results showed that the efficiency was higher in the conditions described as follows: Excised leaves were inoculated with A. tumefaciens for 5~8 min, washed by MS containing 500 mg/L Cef and transferred to co-culture medium for three days, and then placed on selection medium containing 300 mg/L Cef and kan increased from 5 mg/L to 20 mg/L. Seven resistant plants were acquired in transgenic process and detected by PCR, RT-PCR, green fluorescence observation and protein signal analysis. Six of the seven transgenic plants were verified to be positive in PCR detection, green fluorescence and fluorescence protein signal in three of the six plants leaves (T1, T3 and T6) was stronger. The results of the molecular detection and fluorescence detection were the same, so it is possible to find transgenic plant by detecting the green fluorescence directly and achieves the goal for optimizing genetic transformation system for Malus baccata Borkh.
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Received: 29 July 2013
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