title: The intergenic transcribed spacer region 1 as a molecular marker for identification and discrimination of Enterobacteriaceae associated with acute oak decline.
abstract: OBJECTIVE
  We assessed the veracity of intergenic spacer region 1 (ITS1) ribotyping for the rapid, inexpensive and accurate identification of Brenneria goodwinii and Gibbsiella quercinecans that are associated with acute oak decline (AOD) in the UK.
  
  
  RESULTS
  Agarose gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) were applied for the typing of ITS1 PCR amplicons from strains of B. goodwinii, G. quercinecans and related species (n = 34). The number and length of ITS1 amplicons varied significantly between strains. ITS1 profiles generated via PAGE were used to differentiate species using a neighbour-joining phylogram. The ITS1 phylogram was compared against DNA gyrase B (gyrB) gene sequences from the same strains, demonstrating that ITS1 ribotyping is as effective as gyrB at resolving G. quercinecans and B. goodwinii to the species level.
  
  
  CONCLUSIONS
  The ITS1 gene has been successfully employed as a novel marker to resolve newly described AOD-associated Enterobacteriaceae, B. goodwinii and G. quercinecans, to species level.
  
  
  CONCLUSIONS
  ITS1 ribotyping of B. goodwinii and G. quercinecans provides equivalent sensitivity to the current standard method for strain identification (sequence analysis of the gyrB gene), but with reduced processing time and cost. Furthermore, the ITS1 gene is widely applicable as a rapid and inexpensive typing system for Enterobacteriaceae.
authors.affiliation: School of Biological Sciences, Bangor University, Bangor, UK.
authors.forename: J
authors.initials: J
authors.lastname: Doonan
authors.forename: S
authors.initials: S
authors.lastname: Denman
authors.forename: C
authors.initials: C
authors.lastname: Gertler
authors.forename: J A
authors.initials: JA
authors.lastname: Pachebat
authors.forename: P N
authors.initials: PN
authors.lastname: Golyshin
authors.forename: J E
authors.initials: JE
authors.lastname: McDonald
mesh_terms: D027081:DNA Gyrase / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004755:Enterobacteriaceae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D005819:Genetic Markers; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction; D029963:Quercus / Q000382:microbiology*; D021521:Ribotyping / Q000379:methods*
chemical_list: D021903:DNA, Ribosomal Spacer; D005819:Genetic Markers; D027081:DNA Gyrase
keywords: Brenneria goodwinii; DNA gyrase B; Enterobacteriaceae; Gibbsiella quercinecans; ITS1; acute oak decline