Rapid identification of bacteria associated with Acute Oak Decline by high-resolution melt analysis

Brady, C.; Allainguillaume, J.; Denman, S.; Arnold, D.

Abstract

© 2016 The Society for Applied Microbiology Two Gram-negative Enterobacteriaceae, Gibbsiella quercinecans and Brenneria goodwinii, are frequently isolated from oak suffering from Acute Oak Decline. These two species are difficult to identify based on colony morphology, carbohydrate utilization or 16S rRNA gene sequence, and identification using gyrB gene sequencing is time-consuming and laborious. A rapid identification technique, based on high-resolution melt analysis of the atpD gene, was designed to efficiently process numerous isolates from an increasing number of affected woodlands and parks. Principal component analysis of the resulting melt curves from strains of G.quercinecans, B.goodwinii and their close phylogenetic relatives allowed differentiation into distinct clusters based on species or subspecies identity. Significance and Impact of the Study: Acute Oak Decline is an increasing threat to Britain's native oak population. Two novel bacterial species both belonging to the family Enterobacteriaceae, Gibbsiella quercinecans and Brenneria goodwinii, are thought to play an important role in symptom development. Here, we describe a rapid identification technique using high-resolution melt analysis of the atpD gene able to assign isolates to either G.quercinecans or B.goodwinii in a single assay, greatly reducing the time taken to identify if either or both of these species are present in symptomatic oak.

• Journal Article Type: Article
• Acceptance Date: May 23, 2016
• Online Publication Date: May 26, 2016
• Publication Date: Aug 1, 2016
• Deposit Date: May 24, 2016
• Publicly Available Date: May 26, 2017
• Journal: Letters in Applied Microbiology
• Print ISSN: 0266-8254
• Electronic ISSN: 1472-765X
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 63
• Issue: 2
• Pages: 89-95
• DOI: https://doi.org/10.1111/lam.12593
• Keywords: Gibbsiella quercinecans, Brenneria goodwinii, Acute Oak Decline, high resolution melt, HRM, identification
• Public URL: https://uwe-repository.worktribe.com/output/908747
• Publisher URL: http://dx.doi.org/10.1111/lam.12593
• Additional Information: Additional Information : This is the peer reviewed version of the following article: Brady, C., Allainguillaume, J., Denman, S. and Arnold, D. (2016) Rapid identification of bacteria associated with Acute Oak Decline by high resolution melt (HRM) analysis. Letters in Applied Microbiology. 63 (2), pp.89-95, which has been published in final form at http://dx.doi.org/10.1111/lam.12593. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
• Contract Date: May 24, 2016