title: 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.
authors.affiliation: Department of Environment and Primary Industries, Biosciences Research Division, AgriBio, La Trobe University, 5 Ring Road, Bundoora, Victoria 3083, Australia; Plant Biosecurity Cooperative Research Centre, LPO Box 5012, Bruce, ACT, 2617, Australia. Electronic address: linda.zheng@depi.vic.gov.au.
authors.forename: Linda
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Plant Health and Environment Laboratory, Ministry for Primary Industries, P.O. Box 2095, Auckland 1140, New Zealand. Electronic address: joe.tang@mpi.govt.nz.
authors.forename: Joe
authors.initials: J
authors.lastname: Tang
authors.affiliation: Plant Health and Environment Laboratory, Ministry for Primary Industries, P.O. Box 2095, Auckland 1140, New Zealand.
authors.forename: Gerard R G
authors.initials: GR
authors.lastname: Clover
authors.affiliation: Department of Environment and Primary Industries, Grains Innovation Park, 110 Natimuk Rd, Horsham, Victoria 3400, Australia.
authors.forename: Merrin E
authors.initials: ME
authors.lastname: Spackman
authors.affiliation: Plant Biosecurity Cooperative Research Centre, LPO Box 5012, Bruce, ACT, 2617, Australia; Department of Environment and Primary Industries, Grains Innovation Park, 110 Natimuk Rd, Horsham, Victoria 3400, Australia. Electronic address: angela.freeman@depi.vic.gov.au.
authors.forename: Angela J
authors.initials: AJ
authors.lastname: Freeman
authors.affiliation: Department of Environment and Primary Industries, Biosciences Research Division, AgriBio, La Trobe University, 5 Ring Road, Bundoora, Victoria 3083, Australia; Plant Biosecurity Cooperative Research Centre, LPO Box 5012, Bruce, ACT, 2617, Australia. Electronic address: brendan.rodoni@depi.vic.gov.au.
authors.forename: Brendan C
authors.initials: BC
authors.lastname: Rodoni
mesh_terms: D001315:Australia; D017931:DNA Primers / Q000235:genetics*; D002523:Edible Grain / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012328:RNA Viruses / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers
keywords: Area of freedom; Biosecurity; Degenerate primer; Plant virus detection; RT-PCR; Universal primer