Detection of an unidentified potyvirus from Roystonea regia palm using the polymerase chain reaction and degenerate, potyvirus specific, primers and potential problems arising from the amplification of host plant DNA sequences.
Degenerate potyvirus-specific primers were used in the PCR to amplify cDNA representing a 335 nucleotide region of the coat protein gene in RNA purified from an uncharacterised potyvirus isolated from Roystonea regia palms in Queensland, Australia. The RNA was also detected by PCR in total nucleic acid extracts from infected Nicotiana benthamiana, N. clevelandii and Vicia faba. The method also successfully detected pea seed-borne mosaic virus in Pisum sativum. In addition the procedure amplified DNA's of approximately 200 bp and 420 bp, of non-viral origin, from total nucleic acid extracts of healthy Nicotiana spp, indicating that size of the PCR products needs to be included as a criterion for identifying virus specific products when this method is used.
Pearson
M N
MN
School of Biological Sciences, University of Auckland, New Zealand.
Thomas
J E
JE
Randles
J W
JW
D001483:Base Sequence; D017931:DNA Primers; D018744:DNA, Plant / Q000032:analysis*; D008969:Molecular Sequence Data; D010944:Plants / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D017800:Potyvirus / Q000302:isolation & purification*
D017931:DNA Primers; D018744:DNA, Plant