The role of dsbA in colonization of the wheat rhizosphere by Pseudomonas fluorescens Q8r1-96.
Certain well-conserved genes in fluorescent Pseudomonas spp. are involved in pathogenic interactions between the bacteria and evolutionarily diverse hosts including plants, insects and vertebrate animals. One such gene, dsbA, encodes a periplasmic disulfide-bond-forming enzyme implicated in the biogenesis of exported proteins and cell surface structures. This study focused on the role of dsbA in Pseudomonas fluorescens Q8r1-96, a biological control strain that produces the antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) and is known for its exceptional ability to colonize the roots of wheat and pea. The deduced DsbA protein from Q8r1-96 is similar to other predicted thiol : disulfide interchange proteins and contains a conserved DsbA catalytic site, a pattern associated with the thioredoxin family active site, and a signal peptide and cleavage site. A dsbA mutant of Q8r1-96 exhibited decreased motility and fluorescence, and altered colony morphology; however, it produced more 2,4-DAPG and total phloroglucinol-related compounds and was more inhibitory in vitro to the fungal root pathogen Gaeumannomyces graminis var. tritici than was the parental strain. When introduced separately into a natural soil, Q8r1-96 and the dsbA mutant did not differ in their ability to colonize the rhizosphere of wheat in greenhouse experiments lasting 12 weeks. However, when the two strains were co-inoculated, the parental strain consistently out-competed the dsbA mutant. It was concluded that dsbA does not contribute to the exceptional rhizosphere competence of Q8r1-96, although the dsbA mutation reduces competitiveness when the mutant competes with the parental strain in the same niche in the rhizosphere. The results also suggest that exoenzymes and multimeric cell surface structures are unlikely to have a critical role in root colonization by this strain.
Mavrodi
Olga V
OV
Department of Plant Pathology, Washington State University, Pullman, WA, USA.
Mavrodi
Dmitri V
DV
Department of Plant Pathology, Washington State University, Pullman, WA, USA.
Park
Amanda A
AA
USDA-ARS, Root Disease and Biological Control Research Unit, Washington State University, Pullman, WA, USA.
Weller
David M
DM
USDA-ARS, Root Disease and Biological Control Research Unit, Washington State University, Pullman, WA, USA.
Thomashow
Linda S
LS
USDA-ARS, Root Disease and Biological Control Research Unit, Washington State University, Pullman, WA, USA.|Department of Plant Pathology, Washington State University, Pullman, WA, USA.
D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010696:Phloroglucinol / Q000031:analogs & derivatives / Q000378:metabolism; D018517:Plant Roots / Q000382:microbiology*; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism*; D011551:Pseudomonas fluorescens / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014908:Triticum / Q000382:microbiology*
D000900:Anti-Bacterial Agents; C059817:2,4-diacetylphloroglucinol; D010696:Phloroglucinol; D019704:Protein Disulfide-Isomerases