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 PC R in total nucleic acid extracts from infected Nicotiana benthamiana, N. clevelandii and Vicia faba. The method also successfully detected p ea seed-borne mosaic virus in Pisum sativum. In addition the procedure amplified DNA's of approximately 200 bp and 420 bp, of non-viral orig in, from total nucleic acid extracts of healthy Nicotiana spp, indicat ing that size of the PCR products needs to be included as a criterion for identifying virus specific products when this method is used.