Identification of seven novel virulence genes from Xanthomonas citri subsp. citri by Tn5-based random mutagenesis

Xue Song; Jing Guo; Wen-xiu Ma; Zhi-yuan Ji; Li-fang Zou; Gong-you Chen; Hua-song Zou

doi:10.1007/s12275-015-4589-3

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Identification of seven novel virulence genes from Xanthomonas citri subsp. citri by Tn5-based random mutagenesis: Xue Song , Jing Guo , Wen-xiu Ma , Zhi-yuan Ji , Li-fang Zou , Gong-you Chen , Hua-song Zou; Journal of Microbiology 2015;53(5):330-336.
DOI: https://doi.org/10.1007/s12275-015-4589-3
Published online: May 3, 2015

School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, P. R. ChinaSchool of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, P. R. China

Corresponding author: Gong-you Chen , Tel: +86-21-34205873,
Hua-song Zou , Tel: +86-21-34205873,

Received: 16 October 2014 • Revised: 3 March 2015 • Accepted: 28 March 2015

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Abstract

To identify novel virulence genes, a mutant library of Xanthomonas citri subsp. citri 29-1 was produced using EZ-Tn5 transposon and the mutants were inoculated into susceptible grapefruit. Forty mutants with altered virulence phenotypes were identified. Nine of the mutants showed a complete loss of citrus canker induction, and the other 31 mutants resulted in attenuated canker symptoms. Southern blot analysis revealed that each of the mutants carried a single copy of Tn5. The flanking sequence was identified by plasmid rescue and 18 different ORFs were identified in the genome sequence. Of these 18 ORFs, seven had not been previously associated with the virulence of X. citri subsp. citri and were therefore confirmed by complementation analysis. Real-time PCR analysis showed that the seven genes were upregulated when the bacteria were grown in citrus plants, suggesting that the expression of these genes was essential for canker development.

Keywords: Xanthomonas citri subsp. citri;mutant library;virulence genes;canker development

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Identification of seven novel virulence genes from Xanthomonas citri subsp. citri by Tn5-based random mutagenesis

J. Microbiol. 2015;53(5):330-336. Published online May 3, 2015

DOI: https://doi.org/10.1007/s12275-015-4589-3

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