A generic RT‐PCR assay for the detection of Luteoviridae

Plant Pathology - Tập 59 Số 3 - Trang 429-442 - 2010
Anastasija Podolyan1, M. N. Pearson2, G. R. G. Clover3, K. Farreyrol2, David Saul4, J. G. Hampton1, Karen Armstrong1
1Bio-Protection Research Centre, PO Box 84, Lincoln University, Lincoln 7647
2School of Biological Sciences, University of Auckland, PO Box 92019, Auckland
3Plant Health and Environment Laboratory, Investigation and Diagnostic Centre, MAF Biosecurity New Zealand, PO Box 2095, Auckland 1140
4ZyGEM Corporation Ltd, Hamilton NZ, Waikato Innovation Park, Ruakura Road, Hamilton, New Zealand

Tóm tắt

This study, using RT‐PCR, is the first comprehensive assessment since 1991 of a generic detection method for the Luteoviridae. Thirteen Luteoviridae species were detected using three separate sets of low‐degeneracy generic primers with RT‐PCR to amplify 68‐, 75‐ and 129/156‐bp regions of the Luteoviridae coat‐protein gene. Species detected include all members of the genus Luteovirus [Barley yellow dwarf virus (BYDV)‐PAV, BYDV‐PAS, BYDV‐MAV (129 and/or 156 bp amplicons), Soybean dwarf virus, Bean leafroll virus (68 bp amplicon)] and eight of nine species from the genus Polerovirus [Beet western yellows virus, Beet chlorosis virus, Beet mild yellowing virus, Turnip yellows virus, Potato leafroll virus, Cucurbit aphid‐borne yellows virus, Cereal yellow dwarf virus‐RPV (68‐bp amplicon) and Sugarcane yellow leaf virus (75‐bp amplicon)]. These primers were not able to detect Carrot red leaf virus, Sweet potato leaf speckling virus (both belong to unassigned Luteoviridae) and Pea enation mosaic virus‐1 (genus Enamovirus). A synthetic positive control containing all primer sequence priming sites was designed to facilitate this method as a generic tool for use with a variety of host plants. The Luteoviridae primers described in this study present a simple infection‐detection tool of benefit to biosecurity authorities in nursery‐stock surveillance, disease management or outbreak prevention, and may also be useful in detection of as‐yet undiscovered species within the Luteovirus and Polerovirus genera.

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Plant Pathology

Tập 59 Số 3

429-442

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