Novel genus-specific broad range primers for the detection of furoviruses, hordeiviruses and rymoviruses and their application in field surveys in South-east Australia

Abstract

A number of viruses from the genera Furovirus, Hordeivirus and Rymovirus are known to infect and damage the four major temperate cereal crops, wheat, barley, sorghum and oats. Currently, there is no active testing in Australia for any of these viruses, which pose a significant biosecurity threat to the phytosanitary status of Australia's grains industry. To address this, broad spectrum PCR assays were developed to target virus species within the genera Furovirus, Hordeivirus and Rymovirus. Five sets of novel genus-specific primers were designed and tested in reverse-transcription polymerase chain reaction assays against a range of virus isolates in plant virus diagnostic laboratories in both Australia and New Zealand. Three of these assays were then chosen to screen samples in a three-year survey of cereal crops in western Victoria, Australia. Of the 8900 cereal plants screened in the survey, all were tested free of furoviruses, hordeiviruses and rymoviruses. To date, there were no published genus-specific primers available for the detection of furoviruses, hordeiviruses and rymoviruses. This study shows for the first time a broad-spectrum molecular test being used in a survey for exotic grain viruses in Australia. Results from this survey provide important evidence of the use of this method to demonstrate the absence of these viruses in Victoria, Australia. The primer pairs reported here are expected to detect a wide range of virus species within the three genera.

Virus Diseases of Cereal and Oilseed Crops in Australia: Current Position and Future Challenges

“…This is done visually for virus symptoms and by active diagnostic testing for specific viral pathogens as required by Australian federal biosecurity guidelines. After importation requirements are satisfied, healthy planting material can be released [15,[312][313][314]. Should attempts to prevent virus entry and subsequent establishment fail, prompt post border application of eradication or containment procedures is required to avoid further spread [7,8,15,312].…”

The Study of Viral RNA Diversity in Bird Samples Using De Novo Designed Multiplex Genus-Specific Primer Panels

“…This approach has been known for a long time [10] and has been used successfully by various researchers, both in the studies of the representatives of individual viral genera and in the analyses of the diversity of viruses in different types of biological material [11][12][13]. Till recently, a large number of such primers have been described [14][15][16]. However, most of them were designed for the detection of certain species of viruses.…”