Ee. Perez et al., Tobacco rattle virus detection in potato tubers from northeast Florida by PCR and tissue blotting, AM J POTATO, 77(6), 2000, pp. 363-368
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).