Identification and differentiation of Phyllosticta species causing freckle disease of banana using high resolution melting (HRM) analysis

Wong, M. -H., Henderson, J. and Drenth, A. (2013) Identification and differentiation of Phyllosticta species causing freckle disease of banana using high resolution melting (HRM) analysis. Plant Pathology, 62 6: 1285-1293. doi:10.1111/ppa.12056


Author Wong, M. -H.
Henderson, J.
Drenth, A.
Title Identification and differentiation of Phyllosticta species causing freckle disease of banana using high resolution melting (HRM) analysis
Formatted title
Identification and differentiation of Phyllosticta species causing freckle disease of banana using high resolution melting (HRM) analysis
Journal name Plant Pathology   Check publisher's open access policy
ISSN 0032-0862
1365-3059
Publication date 2013-12
Year available 2013
Sub-type Article (original research)
DOI 10.1111/ppa.12056
Volume 62
Issue 6
Start page 1285
End page 1293
Total pages 9
Place of publication West Sussex United Kingdom
Publisher Wiley-Blackwell Publishing
Collection year 2014
Language eng
Formatted abstract
Freckle disease of banana is caused by three closely related species of Phyllosticta, namely P. musarum, P. maculata and P. cavendishii. In this study, a high resolution melting (HRM) analysis assay was developed and its potential to identify these three fungal species is reported. The assay, which targets the ITS of the nuclear rDNA of the fungal species, generates three distinct melt profiles for the three Phyllosticta species. It is also able to distinguish a combination of up to three co-infecting species by generating a deviant melt curve. Thirty-five fungal cultures and infected herbarium leaf specimens, previously characterized using nucleotide sequencing as belonging to one of the three Phyllosticta species, were used for validation of the HRM analysis assay. The normalized curves generated differentiated all samples, with samples from each species correctly identified. The assay was further evaluated against 18 uncharacterized infected leaf specimens from various geographic locations and the results were verified by subsequent nucleotide sequencing. This HRM analysis assay allows rapid identification and differentiation of the three Phyllosticta species using a single primer pair in a one-step closed-tube system without labelled fluorescence probes. This novel assay format has potential for simultaneously identifying and differentiating other closely related species of plant pathogens, as well as the classification of infected historic specimens.
Keyword diagnostics
genotyping
Guignardia
Hrm
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: School of Agriculture and Food Sciences
Queensland Alliance for Agriculture and Food Innovation
Official 2014 Collection
 
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