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Real-Time PCR Assays for the Detection of Puccinia psidii

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Baskarathevan, J., Taylor, R. K., Ho, W., McDougal, R. L., Shivas, R. G. and Alexander, B. J. R. (2016) Real-Time PCR Assays for the Detection of Puccinia psidii. Plant Disease, 100 (3). pp. 617-624.


Article Link: http://dx.doi.org/10.1094/PDIS-08-15-0851-RE


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 β-tubulin gene sequences to detect P. psidii. All qPCR assays were able to detect P. psidii DNA extracted from urediniospores and from infected 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 assay was 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.

Item Type:Article
Business groups:Biosecurity Queensland
Subjects:Plant pests and diseases > Plant pathology
Plant pests and diseases > Pest control and treatment of diseases. Plant protection
Live Archive:01 Jun 2016 06:48
Last Modified:03 Sep 2021 16:44

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