Skip to main content

Research Repository

Advanced Search

The identification of genes important in pseudomonas syringae pv. phaseolicola plant colonisation using in vitro screening of transposon libraries

Neale, Helen C.; Manoharan, Bharani; Neale, Helen C.; Hancock, John T.; Jackson, Robert; Jackson, Robert W.; Arnold, Dawn L.

Authors

Helen C. Neale

Bharani Manoharan

Helen Neale Helen2.Neale@uwe.ac.uk
Research Fellow in Plant Pathology

Robert Jackson

Robert W. Jackson

Dawn Arnold Dawn.Arnold@uwe.ac.uk
Professor in Molecular Plant Pathology



Abstract

© 2015 Manoharan et al. The bacterial plant pathogen Pseudomonas syringae pv. phaseolicola (Pph) colonises the surface of common bean plants before moving into the interior of plant tissue, via wounds and stomata. In the intercellular spaces the pathogen proliferates in the apoplastic fluid and forms microcolonies (biofilms) around plant cells. If the pathogen can suppress the plant's natural resistance response, it will cause halo blight disease. The process of resistance suppression is fairly well understood, but the mechanisms used by the pathogen in colonisation are less clear. We hypothesised that we could apply in vitro genetic screens to look for changes in motility, colony formation, and adhesion, which are proxies for infection, microcolony formation and cell adhesion. We made transposon (Tn) mutant libraries of Pph strains 1448Aand 1302Aand found 106/1920 mutants exhibited alterations in colony morphology, motility and biofilm formation. Identification of the insertion point of the Tn identified within the genome highlighted, as expected, a number of altered motility mutants bearing mutations in genes encoding various parts of the flagellum. Genes involved in nutrient biosynthesis, membrane associated proteins, and a number of conserved hypothetical protein (CHP) genes were also identified. A mutation of one CHP gene caused a positive increase in in planta bacterial growth. This rapid and inexpensive screening method allows the discovery of genes important forin vitro traits that can be correlated to roles in the plant interaction. Copyright:

Journal Article Type Article
Publication Date Sep 1, 2015
Journal PLoS ONE
Electronic ISSN 1932-6203
Publisher Public Library of Science
Peer Reviewed Peer Reviewed
Volume 10
Issue 9
APA6 Citation Jackson, R. W., Neale, H. C., Manoharan, B., Neale, H., Hancock, J. T., Jackson, R., & Arnold, D. L. (2015). The identification of genes important in pseudomonas syringae pv. phaseolicola plant colonisation using in vitro screening of transposon libraries. PLoS ONE, 10(9), https://doi.org/10.1371/journal.pone.0137355
DOI https://doi.org/10.1371/journal.pone.0137355
Keywords genes, transposon libraries
Publisher URL http://dx.doi.org/10.1371/journal.pone.0137355
;