Improved dominant selection markers and co-culturing conditions for efficient Agrobacterium tumefaciens-mediated transformation of Ustilago scitaminea
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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.
KeywordsAgrobacterium tumefaciens-mediated transformation Codon-optimized hygromycin B phosphotransferase gene Recalcitrant transformation Sugar-cane smut disease Ustilago scitaminea
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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