High-quality mutant libraries of Xanthomonas oryzae pv. oryzae and X. campestris pv. campestris generated by an efficient transposon mutagenesis system

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 Tn 5 -based transposome. A highly efficient transformation up to 10 6 transformants per μg transposon DNA was obtained. Southern blot and thermal asymmetric interlaced polymerase chain reaction analyses of Tn 5 insertion sites suggested a random mode of transposition. The transposition was stable in the transformants for 20 subcultures. Eighteen thousand and 17 000 transformants for Xoo and Xcc , respectively, were generated, corresponding to 4 X 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 .