Tobacco rattle virus detection in potato tubers from northeast Florida by PCR and tissue blotting

Tobacco rattle virus (TRV) was detected in potato tubers by reverse transcr iption of RNA followed by amplification of the cDNA by polymerase chain rea ction (PCR). Primer A complementary to residues 6555 to 6575 and primer B i dentical to residues 6113 to 6132 of TRV-SYM RNA-1 were used. After amplifi cation, the 463- bp fragment was cloned, sequenced, and compared to sequenc es of TRV isolates from Europe and Canada. Nucleotide sequence homology of the 16 kDa ORF between the FLA TRV-isolate and the CAN, TCM, PLB, PSG, and SYM TRV-isolates were 94%, 94%, 93%, 90%, and 90%, respectively. A nonradio active biotin-labeled probe detected TRV by tuber tissue blotting. Nematode -transmitted TRV was also detected in bait plants of Nicotiana tabacum cv S amsum, N. clevelandii, and Petunia hybrida by enzyme-linked immunosorbent a ssay (ELISA).