Overexpression of hns in the plant growth-promoting bacterium Enterobacter cloacae UW5 increases root colonization

J Appl Microbiol. 2010 Jun;108(6):2180-90. doi: 10.1111/j.1365-2672.2009.04620.x. Epub 2009 Nov 7.

Abstract

Aims: Plant growth-promoting rhizobacteria (PGPR) introduced into soil often do not compete effectively with indigenous micro-organisms for plant colonization. The aim of this study was to identify novel genes that are important for root colonization by the PGPR Enterobacter cloacae UW5.

Methods and results: A library of transposon mutants of Ent. cloacae UW5 was screened for mutants with altered ability to colonize canola roots using a thermal asymmetric interlaced (TAIL)-PCR-based approach. A PCR fragment from one mutant was reproducibly amplified at greater levels from genomic DNA extracted from mutant pools recovered from seedling roots 6 days after seed inoculation compared to that from the cognate inoculum cultures. Competition assays confirmed that the purified mutant designated Ent. cloacae J28 outcompetes the wild-type strain on roots but not in liquid cultures. In Ent. cloacae J28, the transposon is inserted upstream of the hns gene. Quantitative RT-PCR showed that transposon insertion increased expression of hns on roots.

Conclusions: These results indicate that increased expression of hns in Ent. cloacae enhances competitive colonization of roots.

Significance and impact of the study: A better understanding of the genes involved in plant colonization will contribute to the development of PGPR that can compete more effectively in agricultural soils.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics*
  • Brassica / growth & development
  • Brassica / microbiology*
  • DNA Transposable Elements
  • DNA, Bacterial / genetics
  • DNA-Binding Proteins / genetics*
  • Enterobacter cloacae / genetics*
  • Enterobacter cloacae / growth & development
  • Enterobacter cloacae / metabolism
  • Genes, Bacterial
  • Mutagenesis, Insertional
  • Mutation
  • Plant Roots / growth & development
  • Plant Roots / microbiology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Soil Microbiology*

Substances

  • Bacterial Proteins
  • DNA Transposable Elements
  • DNA, Bacterial
  • DNA-Binding Proteins
  • H-NS protein, bacteria