Production of herbicide-resistant transgenic sweet potato plants through <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> method |
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Authors: | Hye Jin Choi Thummala Chandrasekhar Hyo-Yeon Lee Kyung-Moon Kim |
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Institution: | (1) Kumho Life and Environmental Science Laboratory, Chonnam National University, 1 Oryong-dong, Buk-gu, Gwangju, 500-712, Republic of Korea;(2) Faculty of Biotechnology, Cheju National University, 1 Ara-dong, Jeju, 690-756, Republic of Korea;(3) Subtropical Horticulture Research Center, Cheju National University, 1 Ara-dong, Jeju, 690-756, Republic of Korea |
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Abstract: | Transgenic herbicide-resistant sweet potato plants Ipomoea batatas (L.) Lam.] were produced through Agrobacterium-mediated transformation system. Embryogenic calli derived from shoot apical meristems were infected with Agrobacterium tumefaciens strain EHA105 harboring the pCAMBIA3301 vector containing the bar gene encoding phosphinothricin N-acetyltransferase (PAT) and the gusA gene encoding β-glucuronidase (GUS). The PPT-resistant calli and plants were selected with 5 and 2.5 mg l−1 PPT, respectively. Soil-grown plants were obtained 28–36 weeks after Agrobacterium-mediated transformation. Genetic transformation of the regenerated plants growing under selection was demonstrated by PCR,
and Southern blot analysis revealed that one to three copies of the transgene were integrated into the plant genome of each
transgenic plant. Expression of the bar gene in transgenic plants was confirmed by RT-PCR and application of herbicide. Transgenic plants sprayed with Basta containing
900 mg l−1 of glufosinate ammonium remained green and healthy. The transformation frequency was 2.8% determined by herbicide application
which was high when compared to our previous biolistic method. In addition, possible problems with multiple copies of transgene
were also discussed. We therefore report here a successful and reliable Agrobacterium-mediated transformation of the bar gene conferring herbicide-resistance and this method may be useful for routine transformation and has the potential to develop
new varieties of sweet potato with several important genes for value-added traits such as enhanced tolerance to the herbicide
Basta. |
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Keywords: | Agrobacterium tumefaciens bar Basta gusA Ipomoea batatas Transformation |
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