High-quality mutant libraries of Xanthomonas oryzae pv. oryzae and X. campestris pv. campestris generated by an efficient transposon mutagenesis system |
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Authors: | Sun Qihong Wu Wei Qian Wei Hu Jun Fang Rongxiang He Chaozu |
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Affiliation: | Laboratory of Plant Biotechnology, Institute of Microbiology, The Chinese Academy of Sciences, 100080, Beijing, PR China. |
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Abstract: | A novel transposon mutagenesis system for the phytopathogenic bacteria Xanthomonas oryzae pv. oryzae (Xoo) and X. campestris pv. campestris (Xcc) was developed using a Tn5-based transposome. A highly efficient transformation up to 10(6) transformants per microg transposon DNA was obtained. Southern blot and thermal asymmetric interlaced polymerase chain reaction analyses of Tn5 insertion sites suggested a random mode of transposition. The transposition was stable in the transformants for 20 subcultures. Eighteen thousand and 17000 transformants for Xoo and Xcc, respectively, were generated, corresponding to 4X ORF coverage of the genomes. The libraries will facilitate the identification of pathogenicity-related genes as well as functional genomic analysis in Xoo and Xcc. |
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Keywords: | Xanthomonas oryzae pv. oryzae Xanthomonas campestris pv. campestris Mutant library Tn5-based transposome |
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