Real-time PCR assays for the detection of Puccinia psidii

Baskarathevan, Jeyaseelan and Taylor, R. K. and Ho, W. and Alexander, B. J. R. and McDougal, R. L. and Shivas, R. G. (2016) Real-time PCR assays for the detection of Puccinia psidii. Plant Disease, 100 (3). pp. 617-624. ISSN 0191-2917

Abstract

Puccinia psidii (Myrtle rust) is an emerging pathogen that has a wide host range in the Myrtaceae family; it continues to show an increase in geographic range and is considered to be a significant threat to Myrtaceae plants worldwide. In this study, we describe the development and validation of three novel real-time polymerase reaction (qPCR) assays using ribosomal DNA and b-tubulin gene sequences to detect P. psidii. All qPCR assays were able to detect P. psidii DNA extracted from urediniospores and frominfected plants, including asymptomatic leaf tissues.Depending on the gene target, qPCR was able to detect down to 0.011 pg of P. psidii DNA. The most optimum qPCR assay was shown to be highly specific, repeatable, and reproducible following testing using different qPCR reagents and real-time PCR platforms in different laboratories. In addition, a duplex qPCR assaywas developed to allow coamplification of the cytochrome oxidase gene from host plants for use as an internal PCR control. The most optimum qPCR assay proved to be faster and more sensitive than the previously published nested PCR assay and will be particularly useful for high-throughput testing and to detect P. psidii at the early stages of infection, before the development of sporulating rust pustules.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Files associated with this item cannot be displayed due to copyright restrictions.
Faculty / Department / School: No Faculty
Date Deposited: 18 May 2017 21:44
Last Modified: 18 May 2017 21:44
Fields of Research : 06 Biological Sciences > 0607 Plant Biology > 060702 Plant Cell and Molecular Biology
Socio-Economic Objective: E Expanding Knowledge > 97 Expanding Knowledge > 970106 Expanding Knowledge in the Biological Sciences
Identification Number or DOI: 10.1094/PDIS-08-15-0851-RE
URI: http://eprints.usq.edu.au/id/eprint/31335

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