Sequencing and Analysis of the Pseudomonas fluorescens GcM5-1A Genome: A Pathogen Living in the Surface Coat of Bursaphelenchus xylophilus

PLoS One. 2015 Oct 30;10(10):e0141515. doi: 10.1371/journal.pone.0141515. eCollection 2015.

Abstract

It is known that several bacteria are adherent to the surface coat of pine wood nematode (Bursaphelenchus xylophilus), but their function and role in the pathogenesis of pine wilt disease remains debatable. The Pseudomonas fluorescens GcM5-1A is a bacterium isolated from the surface coat of pine wood nematodes. In previous studies, GcM5-1A was evident in connection with the pathogenicity of pine wilt disease. In this study, we report the de novo sequencing of the GcM5-1A genome. A 600-Mb collection of high-quality reads was obtained and assembled into sequence contigs spanning a 6.01-Mb length. Sequence annotation predicted 5,413 open reading frames, of which 2,988 were homologous to genes in the other four sequenced P. fluorescens isolates (SBW25, WH6, Pf0-1 and Pf-5) and 1,137 were unique to GcM5-1A. Phylogenetic studies and genome comparison revealed that GcM5-1A is more closely related to SBW25 and WH6 isolates than to Pf0-1 and Pf-5 isolates. Towards study of pathogenesis, we identified 79 candidate virulence factors in the genome of GcM5-1A, including the Alg, Fl, Waa gene families, and genes coding the major pathogenic protein fliC. In addition, genes for a complete T3SS system were identified in the genome of GcM5-1A. Such systems have proved to play a critical role in subverting and colonizing the host organisms of many gram-negative pathogenic bacteria. Although the functions of the candidate virulence factors need yet to be deciphered experimentally, the availability of this genome provides a basic platform to obtain informative clues to be addressed in future studies by the pine wilt disease research community.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification
  • Genome, Bacterial*
  • Molecular Sequence Data
  • Phylogeny
  • Pinus
  • Plant Diseases / microbiology
  • Pseudomonas fluorescens / classification
  • Pseudomonas fluorescens / genetics*
  • Pseudomonas fluorescens / isolation & purification
  • Pseudomonas fluorescens / pathogenicity
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Tylenchida / microbiology*
  • Tylenchida / pathogenicity
  • Type III Secretion Systems / genetics
  • Virulence

Substances

  • DNA, Bacterial
  • Type III Secretion Systems

Associated data

  • GENBANK/JJOE00000000

Grants and funding

Funding for this work is provided by the Natural Science Foundation (31070575), the State Forestry Administration, People’s Republic of China (20070430), and program for Innovative Research Team of the Educational Department of China and in Universities of Jiangsu Province. It is also supported by the Priority Academic Program Development (PAPD) program at Nanjing Forestry University.