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Genome Sequencing and Transposon Mutagenesis of Burkholderia seminalis TC3.4.2R3 Identify Genes Contributing to Suppression of Orchid Necrosis Caused by B-gladioli

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  • From a screen of 36 plant-associated strains of Burkholderia spp., we identified 24 strains that suppressed leaf and pseudobulb necrosis of orchid caused by B. gladioli. To gain insights into the mechanisms of disease suppression, we generated a draft genome sequence from one suppressive strain, TC3.4.2R3. The genome is an estimated 7.67 megabases in size, with three replicons, two chromosomes, and the plasmid pC3. Using a combination of multilocus sequence analysis and phylogenomics, we identified TC3.4.2R3 as B. seminalis, a species within the Burkholderia cepacia complex that includes opportunistic human pathogens and environmental strains. We generated and screened a library of 3,840 transposon mutants of strain TC3.4.2R3 on orchid leaves to identify genes contributing to plant disease suppression. Twelve mutants deficient in suppression of leaf necrosis were selected and the transposon insertions were mapped to eight loci. One gene is in a wcb cluster that is related to synthesis of extracellular polysaccharide, a key determinant in bacterial-host interactions in other systems, and the other seven are highly conserved among Burkholderia spp. The fundamental information developed in this study will serve as a resource for future research aiming to identify mechanisms contributing to biological control.
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  • Araújo, W. L., Creason, A. L., Mano, E. T., Camargo-Neves, A. A., Minami, S. N., Chang, J. H., & Loper, J. E. (2016). Genome Sequencing and Transposon Mutagenesis of Burkholderia seminalis TC3. 4.2 R3 Identify Genes Contributing to Suppression of Orchid Necrosis Caused by B. gladioli. Molecular Plant-Microbe Interactions, 29(6), 435-446. doi:10.1094/MPMI-02-16-0047-R
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  • 29
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  • 6
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  • This work was supported by grant from Fundacao de Amparo a Pesquisa do Estado de São Paulo (FAPESP) (2012/24217-6 and 2013/02124-9) and Fundação de Amparo ao Ensino e Pesquisa (FAEP) (University of Mogi das Cruzes). Work in the J. H. Chang lab is supported in part by Agriculture and Food Research Initiative Competitive Grants Program (grant numbers 2011-67019-30192 and 2012-67013-19392) from the United States Department of Agriculture (USDA) National Institute of Food and Agriculture, National Science Foundation (grant number IOS-1021463), and National Institute of General Medical Sciences of the National Institutes of Health under award number RO1GM104977. Work in the J. E. Loper lab was supported in part by Agriculture and Food Research Initiative Competitive grant 2011-67019-30192 from the USDA National Institute of Food and Agriculture.
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