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Improved dominant selection markers and co-culturing conditions for efficient Agrobacterium tumefaciens-mediated transformation of Ustilago scitaminea

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Abstract

Ustilago scitaminea is the causal agent of sugar-cane smut disease. There is, however, no genetic transformation method for it. Here we report the development of an efficient mutagenesis method based on Agrobacterium tumefaciens-mediated transformation. To improve transformation efficiency, a range of conditions, including the codon-usage preference of the selection marker gene, promoters and the culture conditions for transformation were optimized. A strong promoter to drive marker gene expression, optimized codon usage of selection marker gene, controlled water content and pH of co-culture medium were critical factors affecting transformation efficiency. Our findings provide a useful tool for genetic analysis of this important plant pathogen.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Project No. 31271993) and the Temasek Trust and the Economic Development Board, Singapore. The authors express sincere gratitude to Prof. Lian-Hui Zhang, Institute of Molecular and Cell Biology, Singapore for his valuable suggestions and revision of the manuscript.

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Authors and Affiliations

  1. Department of Plant Pathology, South China Agricultural University, Guangzhou, 510642, China

    Longhua Sun, Meixin Yan, Zhaojian Ding, Pinggen Xi, Jinling Liao & Zide Jiang

  2. Biomaterials and Biocatalysts Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore, 117604, Singapore

    Longhua Sun, Yanbin Liu, Minge Du & Lianghui Ji

  3. Guangdong Province Key Laboratory of Microbial Signals and Disease Control, South China Agricultural University, Guangzhou, 510642, China

    Pinggen Xi, Jinling Liao & Zide Jiang

Authors
  1. Longhua Sun
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  2. Meixin Yan
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  3. Zhaojian Ding
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  4. Yanbin Liu
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  5. Minge Du
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  6. Pinggen Xi
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  7. Jinling Liao
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  8. Lianghui Ji
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  9. Zide Jiang
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Correspondence to Zide Jiang.

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Sun, L., Yan, M., Ding, Z. et al. Improved dominant selection markers and co-culturing conditions for efficient Agrobacterium tumefaciens-mediated transformation of Ustilago scitaminea . Biotechnol Lett 36, 1309–1314 (2014). https://doi.org/10.1007/s10529-014-1486-5

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  • DOI: https://doi.org/10.1007/s10529-014-1486-5

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