Development of a multiplex PCR diagnostic assay for the detection of Stagonosporopsis species associated with ray blight of Asteraceae

Vaghefi, Niloofar and Hay, Frank S. and Pethybridge, Sarah J. and Ford, Rebecca and Taylor, Paul W. J. (2016) Development of a multiplex PCR diagnostic assay for the detection of Stagonosporopsis species associated with ray blight of Asteraceae. European Journal of Plant Pathology, 146 (3). pp. 581-595. ISSN 0929-1873

Abstract

Ray blight, a destructive disease of Asteraceae worldwide, is caused by three morphologically similar but phylogenetically distinct species; Stagonosporopsis chrysanthemi, S. inoxydabilis and S. tanaceti. Stagonosporopsis chrysanthemi has been reported as a specific pathogen of chrysanthemum while S. inoxydabilis has been found associated with various Asteraceae. Stagonosporopsis tanaceti has only been reported in Australia, causing substantial crop loss on pyrethrum. All three species were shown to infect and cause disease on in vitro grown pyrethrumplants, hence, S. chrysanthemi and S. inoxydabilis may pose a significant biosecurity threat to the Australian pyrethrum industry. All these Stagonosporopsis species are also Level 2 quarantine pathogens in Europe. Rapid and accurate detection and differentiation of these species is a priority for ray blight management in Australia and in Europe. Accordingly, three species-specific PCR-based assays, targeted to the intergenic spacer of the nuclear ribosomal DNA, were developed. The specificity of each assay was confirmed against 21 Stagonosporopsis spp. as well as 14 pathogenic and saprophytic fungal species commonly found in association with pyrethrum in Australia. The primers were highly sensitive and specific to the target species, detecting down to 4 fg of genomic DNA. These primers were further used in a multiplex PCR to differentiate the presence of the three Stagonosporopsis spp. based on variable sized amplicons in a single reaction.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Published version cannot be displayed due to copyright restrictions.
Faculty / Department / School: No Faculty
Date Deposited: 22 Aug 2017 04:07
Last Modified: 22 Aug 2017 04:07
Uncontrolled Keywords: Chrysanthemi; Chrysanthemum; Inoxydabilis: intergenic spacer; pyrethrum; Ribosomal DNA; Tanaceti
Fields of Research : 06 Biological Sciences > 0607 Plant Biology > 060704 Plant Pathology
10 Technology > 1001 Agricultural Biotechnology > 100101 Agricultural Biotechnology Diagnostics (incl. Biosensors)
Socio-Economic Objective: E Expanding Knowledge > 97 Expanding Knowledge > 970107 Expanding Knowledge in the Agricultural and Veterinary Sciences
Identification Number or DOI: 10.1007/s10658-016-0944-4
URI: http://eprints.usq.edu.au/id/eprint/32948

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