Real-time PCR assays for the detection of Puccinia psidii
Article
Article Title | Real-time PCR assays for the detection of Puccinia psidii |
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ERA Journal ID | 2647 |
Article Category | Article |
Authors | Baskarathevan, Jeyaseelan (Author), Taylor, R. K. (Author), Ho, W. (Author), Alexander, B. J. R. (Author), McDougal, R. L. (Author) and Shivas, R. G. (Author) |
Journal Title | Plant Disease: an international journal of applied plant pathology |
Journal Citation | 100 (3), pp. 617-624 |
Number of Pages | 7 |
Year | 2016 |
Publisher | American Phytopathological Society |
Place of Publication | St. Paul, United States |
ISSN | 0191-2917 |
1943-7692 | |
Digital Object Identifier (DOI) | https://doi.org/10.1094/PDIS-08-15-0851-RE |
Web Address (URL) | http://apsjournals.apsnet.org/doi/10.1094/PDIS-08-15-0851-RE |
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. |
ANZSRC Field of Research 2020 | 310803. Plant cell and molecular biology |
Public Notes | Files associated with this item cannot be displayed due to copyright restrictions. |
Byline Affiliations | Ministry for Primary Industries, New Zealand |
Scion, New Zealand | |
Department of Agriculture, Fisheries and Forestry, Queensland | |
Institution of Origin | University of Southern Queensland |
https://research.usq.edu.au/item/q4004/real-time-pcr-assays-for-the-detection-of-puccinia-psidii
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