Efficient <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation using embryogenic suspension cultures in sweetpotato, <Emphasis Type="Italic">Ipomoea batatas</Emphasis> (L.) Lam. |
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Authors: | Bo Yu Hong Zhai Yuping Wang Ning Zang Shaozhen He Qingchang Liu |
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Institution: | (1) Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing, 100094, China;(2) Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing, 100094, China;(3) Laboratory of Crop Heterosis and Utilization, Ministry of Education, China Agricultural University, Beijing, 100094, China |
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Abstract: | Efficient Agrobacterium tumefaciens-mediated transformation was achieved using embryogenic suspension cultures of sweetpotato (Ipomoea batatas (L.) Lam.) cv. Lizixiang. Cell aggregates from embryogenic suspension cultures were cocultivated with the A. tumefaciens strain EHA105 harboring a binary vector pCAMBIA1301 with gusA and hygromycin phosphotransferase II gene (hpt II) genes. Selection culture was conducted using 25 mg l−1 hygromycin. A total of 2,218 plants were regenerated from the inoculated 1,776 cell aggregates via somatic embryogenesis.
β-glucuronidase (GUS) assay and PCR, dot blot and Southern blot analyses of the regenerated plants randomly sampled showed
that 90.37% of the regenerated plants were transgenic plants. The number of integrated T-DNA copies varied from 1 to 4. Transgenic
plants, when transferred to soil in a greenhouse and a field, showed 100% survival. No morphological variations were observed
in the ex vitro transgenic plants. These results exceed all transformation experiments reported so far in the literature in
quantity of independent events per transformation experiment in sweetpotato. |
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Keywords: | Agrobacterium tumefaciens Embryogenic cell suspension Genetic transformation Sweetpotato Transgenic plant |
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