Transposon mutagenesis of Pseudomonas syringae pathovars syringae and morsprunorum to identify genes involved in bacterial canker disease of cherry

Helen C. Neale, Michelle Hulin, Richard Harrison, Robert Jackson, Dawn Arnold

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Abstract

Bacterial canker of Prunus, affecting economically important stone fruit crops including cherry, peach, apricot and plum, is caused by the plant pathogen Pseudomonas syringae (P.s.). Strains from two pathovars—P.s. pv. syringae (Pss) and P.s. pv. morsprunorum race 1 (PsmR1) and 2 (PsmR2)—in three phylogenetically distant clades have convergently evolved to infect Prunus. The bacteria enter woody tissues through wounds and leaf scars, causing black necrotic cankers. Symptoms are also produced on blossom, fruit and leaves. Little is known about the mechanisms P.s. uses to colonise tree hosts such as Prunus. Here, we created transposon (Tn) mutant libraries in one strain of P.s. from each of the three clades and screened the mutants on immature cherry fruit to look for changes in virulence. Mutants (242) with either reduced or enhanced virulence were detected and further characterised by in vitro screens for biofilm formation, swarming ability, and pathogenicity on leaves and cut shoots. In total, 18 genes affecting virulence were selected, and these were involved in diverse functions including motility, type III secretion, membrane transport, amino acid synthesis, DNA repair and primary metabolism. Interestingly, mutation of the effector gene, hopAU1, led to an increase in virulence of Psm R2.
Original languageEnglish
Article number1328
JournalMicroorganisms
Volume9
Issue number6
Early online date18 Jun 2021
DOIs
Publication statusPublished - 18 Jun 2021

Bibliographical note

Funding Information:
This research was funded by the Biotechnology and Biological Sciences Research Council, United Kingdom (BB/P005705/1 and EMR grant number BB/ P006272/1).

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

Keywords

  • Pseudomonas syringae
  • bacterial canker
  • pathogenicity
  • transposon mutagenesis

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