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Biotechnology Letters

, Volume 36, Issue 6, pp 1309–1314 | Cite as

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

  • Longhua Sun
  • Meixin Yan
  • Zhaojian Ding
  • Yanbin Liu
  • Minge Du
  • Pinggen Xi
  • Jinling Liao
  • Lianghui Ji
  • Zide Jiang
Original Research Paper

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.

Keywords

Agrobacterium tumefaciens-mediated transformation Codon-optimized hygromycin B phosphotransferase gene Recalcitrant transformation Sugar-cane smut disease Ustilago scitaminea 

Notes

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.

Supplementary material

10529_2014_1486_MOESM1_ESM.doc (42 kb)
Supplementary material 1 (DOC 43 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Longhua Sun
    • 1
    • 2
  • Meixin Yan
    • 1
  • Zhaojian Ding
    • 1
  • Yanbin Liu
    • 2
  • Minge Du
    • 2
  • Pinggen Xi
    • 1
    • 3
  • Jinling Liao
    • 1
    • 3
  • Lianghui Ji
    • 2
  • Zide Jiang
    • 1
    • 3
  1. 1.Department of Plant PathologySouth China Agricultural UniversityGuangzhouChina
  2. 2.Biomaterials and Biocatalysts Group, Temasek Life Sciences Laboratory, 1 Research LinkNational University of SingaporeSingaporeSingapore
  3. 3.Guangdong Province Key Laboratory of Microbial Signals and Disease ControlSouth China Agricultural UniversityGuangzhouChina

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