The detection of recombinant, tuber necrosing isolates of Potato virus Y (PVY(NTN)) using a three-primer PCR based in the coat protein gene
Language English Country Netherlands Media print
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
12668269
DOI
10.1016/s0166-0934(03)00047-8
PII: S0166093403000478
Knihovny.cz E-resources
- MeSH
- DNA Primers * MeSH
- DNA-Directed RNA Polymerases MeSH
- Molecular Sequence Data MeSH
- Plant Diseases virology MeSH
- Polymerase Chain Reaction methods MeSH
- Polymorphism, Genetic MeSH
- Potyvirus genetics isolation & purification MeSH
- Recombination, Genetic * MeSH
- Base Sequence MeSH
- Sequence Analysis, DNA MeSH
- Solanum tuberosum virology MeSH
- Capsid Proteins genetics MeSH
- Viral Proteins genetics MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA Primers * MeSH
- DNA-Directed RNA Polymerases MeSH
- nuclear inclusion protein b, mosaic viruses MeSH Browser
- Capsid Proteins MeSH
- Viral Proteins MeSH
A simple and reliable procedure for reverse transcription-polymerase chain reaction (RT-PCR) detection and strain differentiation of Potato virus Y (PVY) was developed. Three primers were designed within the coat protein (CP) and nuclear inclusion protein b (NIb) region, exploiting a single base polymorphism identified as being present in all the recombinant PVY(NTN) isolates published. Samples infected with PVY produce a single band of 569 bp, while isolates belonging to PVY(NTN) strain give an additional band of 334 bp. The technique was tested on a collection of well-characterised isolates of PVY from a range of strains and was found to detect all of the isolates reported as belonging to the PVY(NTN) strain. All of the isolates detected possess a recombination event within the coat protein. Further sequence analysis revealed that all the recombinant PVY(NTN) isolates reported thus far would be detected using this assay, whilst isolates thought to be PVY(NTN) that do not possess the coat protein recombination event would not be detected.
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