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Doonan, James
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article
The intergenic transcribed spacer region 1 as a molecular marker for identification and discrimination of Enterobacteriaceae associated with acute oak decline
Abstract
Aims<br/>We assessed the veracity of intergenic spacer region 1 (ITS1) ribotyping for the rapid, inexpensive and accurate identification ofBrenneria goodwinii and Gibbsiella quercinecans that are associated with Acute Oak Decline (AOD) in the UK.<br/><br/>Methods and Results<br/>Agarose gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) were applied for the typing of ITS1 PCR amplicons from strains of Brenneria goodwinii, Gibbsiella quercinecans and related species (n = 34). The number and lengt<br/>h 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. qurerciecans and B. goodwinii to the species level.<br/><br/>Conclusions<br/>The ITS1 gene has been successfully employed as a novel marker to resolve newly described AOD associated Enterobacteriaceae, Brenneria goodwinii<br/>and Gibbsiella quercinecans, to species level.Significance and Impact of Study<br/>ITS1 ribotyping of Brenneria goodwinii and Gibbsiella quercinecans provides equivalent sensitivity to the current standard method for strain identification<br/>(sequence analysis of the gyrB gene), but with reduced processing time and cost.<br/>Furthermore, the ITS1 gene is widely applicable as a rapid and inexpensive typing system for Enterobacteriaceae.