Chemical or Genetic Alteration of Proton Motive Force Results in Loss of Virulence of Burkholderia glumae, the Cause of Rice Bacterial Panicle Blight

Appl Environ Microbiol. 2021 Aug 26;87(18):e0091521. doi: 10.1128/AEM.00915-21. Epub 2021 Aug 26.

Abstract

Rice is an important source of food for more than half of the world's population. Bacterial panicle blight (BPB) is a disease of rice characterized by grain discoloration or sheath rot caused mainly by Burkholderia glumae. B. glumae synthesizes toxoflavin, an essential virulence factor that is required for symptoms of the disease. The products of the tox operons, ToxABCDE and ToxFGHI, are responsible for the synthesis and the proton motive force (PMF)-dependent secretion of toxoflavin, respectively. The DedA family is a highly conserved membrane protein family found in most bacterial genomes that likely function as membrane transporters. Our previous work has demonstrated that absence of certain DedA family members results in pleiotropic effects, impacting multiple pathways that are energized by PMF. We have demonstrated that a member of the DedA family from Burkholderia thailandensis, named DbcA, is required for the extreme polymyxin resistance observed in this organism. B. glumae encodes a homolog of DbcA with 73% amino acid identity to Burkholderia thailandensis DbcA. Here, we created and characterized a B. glumae ΔdbcA strain. In addition to polymyxin sensitivity, the B. glumae ΔdbcA strain is compromised for virulence in several BPB infection models and secretes only low amounts of toxoflavin (∼15% of wild-type levels). Changes in membrane potential in the B. glumae ΔdbcA strain were reproduced in the wild-type strain by the addition of subinhibitory concentrations of sodium bicarbonate, previously demonstrated to cause disruption of PMF. Sodium bicarbonate inhibited B. glumae virulence in rice, suggesting a possible non-toxic chemical intervention for bacterial panicle blight. IMPORTANCE Bacterial panicle blight (BPB) is a disease of rice characterized by grain discoloration or sheath rot caused mainly by Burkholderia glumae. The DedA family is a highly conserved membrane protein family found in most bacterial genomes that likely function as membrane transporters. Here, we constructed a B. glumae mutant with a deletion in a DedA family member named dbcA and report a loss of virulence in models of BPB. Physiological analysis of the mutant shows that the proton motive force is disrupted, leading to reduction of secretion of the essential virulence factor toxoflavin. The mutant phenotypes are reproduced in the virulent wild-type strain without an effect on growth using sodium bicarbonate, a nontoxic buffer that has been reported to disrupt the PMF. The results presented here suggest that bicarbonate may be an effective antivirulence agent capable of controlling BPB without imposing an undue burden on the environment.

Keywords: antibiotic resistance; bacterial panicle blight; membrane protein; plant pathogens; proton motive force; rice; virulence.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • Burkholderia* / drug effects
  • Burkholderia* / genetics
  • Burkholderia* / metabolism
  • Burkholderia* / pathogenicity
  • Carbonyl Cyanide m-Chlorophenyl Hydrazone / pharmacology
  • Onions / microbiology
  • Oryza / microbiology*
  • Plant Diseases / microbiology*
  • Proton-Motive Force*
  • Pyrimidinones / metabolism
  • Sodium Bicarbonate / pharmacology
  • Triazines / metabolism
  • Virulence
  • Virulence Factors / metabolism

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Pyrimidinones
  • Triazines
  • Virulence Factors
  • Carbonyl Cyanide m-Chlorophenyl Hydrazone
  • toxoflavin
  • Sodium Bicarbonate

Supplementary concepts

  • Burkholderia glumae