Improved Dominant Selection Markers and Co-Culturing Conditions for Efficient Agrobacterium Tumefaciens-Mediated Transformation of Ustilago Scitaminea
Biotechnology letters(2014)
摘要
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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关键词
Agrobacterium tumefaciens-mediated transformation,Codon-optimized hygromycin B phosphotransferase gene,Recalcitrant transformation,Sugar-cane smut disease,Ustilago scitaminea
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