An RT-PCR primer pair for the detection of Pospiviroid and its application in surveying ornamental plants for viroids

A primer pair for reverse transcription-polymerase chain reaction (RT-PCR), based on the conserved sequences of the members of genus Pospiviroid was designed to yield a fragment of about 200 base pairs (bp). Since pospiviroids infect a large number of plants species and a few members of the genus Po...

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Bibliographic Details
Published in:Journal of virological methods 2004-03, Vol.116 (2), p.189-193
Main Authors: Bostan, Hidayet, Nie, Xianzhou, Singh, Rudra P
Format: Article
Language:English
Subjects:
Base Sequence
Biological and medical sciences
CEVd
CLVd
Coleus - virology
CSVd
DNA Primers
Fundamental and applied biological sciences. Psychology
Impatiens
IrVd
Microbiology
Plant Diseases - virology
Plants - virology
potato spindle tuber virus
Reverse Transcriptase Polymerase Chain Reaction - methods
Techniques used in virology
tomato chlorotic dwarf virus
Verbena
Vinca
Viroids
Viroids - genetics
Viroids - isolation & purification
Viroids - pathogenicity
Virology
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Summary:A primer pair for reverse transcription-polymerase chain reaction (RT-PCR), based on the conserved sequences of the members of genus Pospiviroid was designed to yield a fragment of about 200 base pairs (bp). Since pospiviroids infect a large number of plants species and a few members of the genus Pospiviroid have been already detected in some ornamental plants, the primer pair was evaluated for its efficacy using ornamental plants. The method of return-polyacrylamide gel electrophoresis (R-PAGE) was used to determine the general presence of viroids in the test samples. Efficacy of the primer pair for members of genus Pospiviroid was demonstrated by the detection of Potato spindle tuber viroid (PSTVd) and Tomato chlorotic dwarf viroid (TCDVd) in potato, Chrysanthemum stunt viroid and Iresine viroid in Verbena and Vinca species, and Citrus exocortis viroid in Impatiens species. Specificity of the primer pair became evident, where additional viroids were detected by R-PAGE in Coleus and Magilla species, but they were not amplified by the Pospiviroid primer. This primer pair would be of benefit in indexing ornamental plants in quarantine samples or in viroid-free certification schemes, irrespective of their actual identity.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2003.11.014