Rapid, large-scale generation of Ds transposant lines and analysis of the Ds insertion sites in rice

Authors

  • Chul Min Kim,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Hai Long Piao,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Soon Ju Park,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Nam Soo Chon,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Byoung Il Je,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Bingyao Sun,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
    2. Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea,
    3. College of Life Sciences, Suzhou University, Suzhou 215006, China,
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  • Sung Han Park,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Jin Young Park,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Eun Jin Lee,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Min Jung Kim,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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  • Woo Sik Chung,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
    2. Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea,
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  • Kon Ho Lee,

    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
    2. Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea,
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  • Young Suk Lee,

    1. Department of Applied Biology and Environmental Sciences, Gyeongsang National University, Jinju 660-701, Korea,
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  • Jeung Joo Lee,

    1. Department of Applied Biology and Environmental Sciences, Gyeongsang National University, Jinju 660-701, Korea,
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  • Yong Jae Won,

    1. Rice Division, Yeongnam Agricultural Research Institute, Milyang 627-803, Korea, and
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  • GiHwan Yi,

    1. Rice Division, Yeongnam Agricultural Research Institute, Milyang 627-803, Korea, and
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  • Min Hee Nam,

    1. Rice Division, Yeongnam Agricultural Research Institute, Milyang 627-803, Korea, and
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  • Young Soon Cha,

    1. Rice Functional Genomics Team, National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea
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  • Doh Won Yun,

    1. Rice Functional Genomics Team, National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea
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  • Moo Young Eun,

    1. Rice Functional Genomics Team, National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea
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  • Chang-deok Han

    Corresponding author
    1. Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea,
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For correspondence (fax +82 55 759 9363; e-mail cdhan@nongae.gsnu.ac.kr).

The first six authors contributed equally to the paper.

Summary

Rapid, large-scale generation of a Ds transposant population was achieved using a regeneration procedure involving tissue culture of seed-derived calli carrying Ac and inactive Ds elements. In the F2 progeny from genetic crosses between the same Ds and Ac starter lines, most of the crosses produced an independent germinal transposition frequency of 10–20%. Also, many Ds elements underwent immobilization even though Ac was expressed. By comparison, in a callus-derived regenerated population, over 70% of plants carried independent Ds insertions, indicating transposition early in callus formation. In the remaining population, the majority of plants carried only Ac. Most of the new Ds insertions were stably transmitted to a subsequent generation. An exceptionally high proportion of independent transposants in the regenerated population means that selection markers for transposed Ds and continual monitoring of Ac/Ds activities may not necessarily be required. By analyzing 1297 Ds-flanking DNA sequences, a genetic map of 1072 Ds insertion sites was developed. The map showed that Ds elements were transposed onto all of the rice chromosomes, with preference not only near donor sites (36%) but also on certain physically unlinked arms. Populations from both genetic crossing and tissue culture showed the same distribution patterns of Ds insertion sites. The information of these mapped Ds insertion sites was deposited in GenBank. Among them, 55% of Ds elements were on predicted open-reading frame (ORF) regions. Thus, we propose an optimal strategy for the rapid generation of a large population of Ds transposants in rice.

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