Colinet, D. and Kummert, J. and Duterme, O. and Lepoivre, P. (1995) Detection of plant viruses by PCR using group or virus specific primers (potyviruses, bymoviruses). EPPO Bulletin, 25 (1-2). pp. 289-299.
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Abstract
A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) utilizing degenerate primers derived from conserved regions in the genome of potyviruses was designed to amplify the 5′ terminal part of the coat protein cistron, which can differ greatly in length between distinct potyviruses. Amplification on total RNA extracted from one sweet potato clone from China yielded 3 fragments of different lengths, and sequence analysis of these fragments indicated mixed infections by 3 distinct potyviruses in the sweet potato clone investigated. Two degenerate primers were also designed to amplify the 5′ terminal part of the coat protein cistron of either potyviruses or bymoviruses. A 3rd degenerate primer was selected from a region conserved in potyviruses and bymoviruses and used with primers specific for 2 different bymoviruses, barley yellow mosaic bymovirus (BaYMV) and barley mild mottle bymovirus (BaMMV). The primers allowed the identification of BaYMV and BaMMV barley plants from the field with single or mixed infection and were used to clarify the aetiology of a viral agent inducing mosaic symptoms on several barley cultivars. It is concluded that amplification of fragments, including the variable 5′ terminal region of the coat protein cistron provides a convenient method for the detection and characterization of potyviruses and bymoviruses without fastidious preliminary work of separation and purification of the components of viral complexes.
| Item Type: | Article |
|---|---|
| Additional Information: | This work was financially supported by the EU (project STD3 N )TS3-CT91-0013) and by IRSIA, Brussels (convention D 1/4-12261/5479a). |
| Uncontrolled Keywords: | Potyviruses, chain reaction, bymoviruses, barley, aetiology. |
| Author Affiliation: | Laboratoire de Pathologie Végétale, Faculté des Sciences Agronomiques, 13 av. Maréchal Juin, 5030 Gembloux, Belgium. |
| Subjects: | Plant Protection Crop Improvement |
| Divisions: | General |
| Depositing User: | Mr Balakrishna Garadasu |
| Date Deposited: | 10 Mar 2013 11:15 |
| Last Modified: | 10 Mar 2013 11:15 |
| Official URL: | http://dx.doi.org/10.1111/j.1365-2338.1995.tb01470... |
| URI: | http://eprints.icrisat.ac.in/id/eprint/9801 |
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