Detection of beet yellows virus by RT-PCR and immunocapture RT-PCR in Tetragonia expansa and Beta vulgaris.
Two sensitive methods, RT-PCR with phenol-extracted RNA or Triton X-100-released RNA and immunocapture RT-PCR (IR-RT-PCR) were used for the detection of Beet yellows virus (BYV) in young and old leaves of Tetragonia expansa and sugar beet (Beta vulgaris) and in sugar beet roots. Four oligonucleotide primer pairs proved suitable for the detection of BYV. The release of BYV RNA with Triton X-100 was shown to be a very effective and easy as compared to isolation of total RNA by phenol extraction with the same or higher sensitivity of subsequent PCR. Using the Triton X-100 release of RNA and IC-RT-PCR the sensitivity of detection was so high that pg amounts of BYV RNA occurring in dilutions up to 10(-6) of saps from young Tetragonia and sugar beet leaves could be detected.
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Department of Plant Protection, Czech University of Agriculture, 165 21 Prague 6, Czech Republic. jiban@vurv.cz
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D035722:Aizoaceae / Q000821:virology*; D027461:Beta vulgaris / Q000821:virology*; D017790:Closterovirus / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D017830:Octoxynol; D019800:Phenol; D018515:Plant Leaves / Q000821:virology; D018517:Plant Roots / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D017931:DNA Primers; D012367:RNA, Viral; D019800:Phenol; D017830:Octoxynol