Mannose selection system used for cucumber transformation |
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Authors: | Zhengquan He ZhenZhen Duan Wei Liang Faju Chen Wei Yao Hongwei Liang Chaoyin Yue Zongxiu Sun Fan Chen Jianwu Dai |
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Institution: | (1) Biotechnology Research Center, China Three Gorges University, Yichang, 443002, P.R. China;(2) Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, P.R. China;(3) National Key Laboratory for Rice Biology, China National Rice Research Institute, Hangzhou, 310006, P.R. China |
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Abstract: | The selectable marker system, which utilizes the pmi gene encoding for phosphomannose-isomerase that converts mannose-6-phosphate to fructose-6-phosphate, was adapted for Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.). Only transformed cells were capable of utilizing mannose as a carbon source. The highest transformation frequency of 23% was obtained with 10 g/l mannose and 10 g/l sucrose in the medium. Molecular, genetic analysis, and PMI activity assay showed that the regenerated shoots contained the pmi gene and the gene was transmitted to the progeny in a Mendelian fashion. The results indicated that the mannose selection system, which is devoid of the disadvantages of antibiotic or herbicide selection, could be used for cucumber Agrobacterium-mediated transformation. |
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Keywords: | Agrobacterium-mediated transformation Cucumis sativus L Phosphomannose-isomerase Positive selection |
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