The importance of cotton disease surveys in Queensland for monitoring endemic diseases and detecting new pathogens and pests

The cotton industry in Australia funds biannual disease surveys conducted by plant pathologists. The objective of these surveys is to monitor the distribution and importance of key endemic pests and record the presence or absence of new or exotic diseases. Surveys have been conducted in Queensland since 2002/03, with surveillance undertaken by experienced plant pathologists. Monitoring of endemic diseases indicates the impact of farming practices on disease incidence and severity. The information collected gives direction to cotton disease research. Routine diagnostics has provided early detection of new disease problems which include 1) the identification of Nematospora coryli, a pathogenic yeast associated with seed and internal boll rot; and 2) Rotylenchulus reniformis, a plant-parasitic nematode. This finding established the need for an intensive survey of the Theodore district revealing that reniform was prevalent across the district at populations causing up to 30% yield loss. Surveys have identified an exotic defoliating strain (VCG 1A) and non-defoliating strains of Verticillium dahliae, which cause Verticillium wilt. An intensive study of the diversity of V. dahliae and the impact these strains have on cotton are underway. Results demonstrate the necessity of general multi-pest surveillance systems in broad acre agriculture in providing (1) an ongoing evaluation of current integrated disease management practices and (2) early detection for a suite of exotic pests and previously unknown pests.

Real-Time PCR Assays for the Detection of Puccinia psidii

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.