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Selectionof nptll transgenic sweetpotato plants using G418 and paromomycin

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Abstract

We have used two aminoglycosides, G418 and paromomycin, to develop a reliable selection system fornptll transgenic sweet-potato (Ipomoea batatas (L.) Lam.). Embryogenic calli derived from shoot apical meristems were bombarded with gold particles coated with pCAMBIA2301, which contained thenptll andgusA genes. When compared on a kill curve that was based on calli proliferation and cell viability, G413-selection proved to be more efficient and had fewer escapes than kanamycin. These bombarded expiants were then selected on G418-containing media. The total time required from bombardment to plant establishment in soil was seven to nine months. Multiple copies of the transgene were integrated into the sweetpotato genome. Northern analysis confirmed transgene expression in the regenerated plants, and a paromomycin assay demonstrated that thenptll gene was functionally expressed in transformed sweetpotato. These molecular analyses and assays all showed that selection with G418 and paromomycin is reliable. So far, we have produced 69 transgenic events with this system, at a transformation frequency of approx. 1.1%. That efficiency is based on the number of transgenic plants obtained and the amount of calli bombarded. Thus, this selection method that combines G418 with paromomycin is now available for selectingnptll transgenic sweetpotato.

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Authors and Affiliations

  1. Kumho Life and Environmental Science Laboratory, Chonnam National University, 500-712, Cwangju, Korea

    Young-Mi Shin, Goh Choe, Byoungchul Shin, Gibum Yi, Kiyoung Yang & Kyung-Moon Kim

  2. Jeju-Regional Innovation Agency, Jeju Hi-Tech Industry Development Institute, 690-121, Jeju, Korea

    Pil-Yong Yun

  3. Mokpo Experiment Station, National Institute of Crop Science RDA, 534-833, Mokpo, Korea

    Joon Seol Lee

  4. Laboratory of Environmental Biotechnology, KRIBB, 305-806, Daejeon, Korea

    Sang-Soo Kwak

  5. Agricultural Plant Stress Research Center, Chonnam National University, 500-757, Gwangju, Korea

    Kyung-Moon Kim

  6. Subtropical Horticulture Research Center, Cheju National University, 690-756, Jeju, Korea

    Kyung-Moon Kim

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  1. Young-Mi Shin
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  2. Goh Choe
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  3. Byoungchul Shin
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  4. Gibum Yi
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  7. Joon Seol Lee
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  9. Kyung-Moon Kim
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Correspondence to Kyung-Moon Kim.

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Shin, YM., Choe, G., Shin, B. et al. Selectionof nptll transgenic sweetpotato plants using G418 and paromomycin. J. Plant Biol. 50, 206–212 (2007). https://doi.org/10.1007/BF03030631

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  • DOI: https://doi.org/10.1007/BF03030631

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