• Open Access

A novel light-dependent selection marker system in plants

Authors

  • Serry Koh,

    1. Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
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  • Hongsup Kim,

    1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea
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  • Jinwoo Kim,

    1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea
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    • Current address: Department of Applied Biology, Gyeongsang National University, Jinju 660-701, Korea.

  • Eunhye Goo,

    1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea
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  • Yun-Jung Kim,

    1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea
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  • Okhee Choi,

    1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea
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  • Nam-Soo Jwa,

    1. Department of Molecular Biology, Sejong University, Seoul, Korea
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  • Jun Ma,

    1. Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry, and Pharmaceutical Science, Okayama University, Okayama, Japan
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  • Tomohisa Nagamatsu,

    1. Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry, and Pharmaceutical Science, Okayama University, Okayama, Japan
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  • Jae Sun Moon,

    Corresponding author
    1. Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
      (fax +82 2 873 2317; email ingyu@snu.ac.kr or jsmoon@kribb.re.kr)
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  • Ingyu Hwang

    Corresponding author
    1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea
      (fax +82 2 873 2317; email ingyu@snu.ac.kr or jsmoon@kribb.re.kr)
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  • tflA (GenBank accession number GQ921834).

(fax +82 2 873 2317; email ingyu@snu.ac.kr or jsmoon@kribb.re.kr)

Summary

Photosensitizers are common in nature and play diverse roles as defense compounds and pathogenicity determinants and as important molecules in many biological processes. Toxoflavin, a photosensitizer produced by Burkholderia glumae, has been implicated as an essential virulence factor causing bacterial rice grain rot. Toxoflavin produces superoxide and H2O2 during redox cycles under oxygen and light, and these reactive oxygen species cause phytotoxic effects. To utilize toxoflavin as a selection agent in plant transformation, we identified a gene, tflA, which encodes a toxoflavin-degrading enzyme in the Paenibacillus polymyxa JH2 strain. TflA was estimated as 24.56 kDa in size based on the amino acid sequence and is similar to a ring-cleavage extradiol dioxygenase in the Exiguobacterium sp. 255-15; however, unlike other extradiol dioxygenases, Mn2+and dithiothreitol were required for toxoflavin degradation by TflA. Here, our results suggested toxoflavin is a photosensitizer and its degradation by TflA serves as a light-dependent selection marker system in diverse plant species. We examined the efficiencies of two different plant selection systems, toxoflavin/tflA and hygromycin/hygromycin phosphotransferase (hpt) in both rice and Arabidopsis. The toxoflavin/tflA selection was more remarkable than hygromycin/hpt selection in the high-density screening of transgenic Arabidopsis seeds. Based on these results, we propose the toxoflavin/tflA selection system, which is based on the degradation of the photosensitizer, provides a new robust nonantibiotic selection marker system for diverse plants.

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