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Identification Of Distinct Potyviruses In Mixedly-Infected Sweet-Potato By The Polymerase Chain-Reaction With Degenerate Primers

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  • Life Sciences :: Phytobiology (Plant Sciences
  • Forestry
  • Mycology...) [F12]
  • Sciences Du Vivant :: Biologie Végétale (Sciences Végétales
  • Sylviculture
  • Mycologie...) [F12]
  • Biology
  • Design


Identification of Distinct Potyviruses in Mixedly-Infected Sweetpotato by the Polymerase Chain Reaction with Degenerate Primers D. Colinet, J. Kummert, P. Lepoivre, and J. Semal Faculté des Sciences Agronomiques de I'Etat, Laboratoire de Pathologie Végétale, 5030 Gembloux, Belgium. Financially supported by EEC (Project STD3 TS3-CT9l-0013). We thank Dr. Kettmann for helpful discussions, Pr. Burny in whose laboratory part of this work was carried out, and Pr. Brunt (AFRC Institute of Horticultural Research, Littlehampton, England) for supplying the SPLV isolate. Accepted for publication 24 September 1993. ABSTRACT Colinet, D., Kummert, J., Lepoivre, P., and Semal, J. 1994. Identification of distinct potyviruses in mixedly-infected sweetpotato by the polymerase chain reaction with degenerate primers. 84:65-69. A combined assay of reverse transcription and the polymerase chain reaction utilizing degenerate primers derived from conserved regions in the genome of potyviruses was designed to amplify the variable 5'-terminal region of the coat protein cistron. Amplification on total RNA extracted from two sweetpotato clones originating from China (Guangdong Province) yielded a I .35-kb fragment amplified from a sweetpotato leathery mottle virus (SPFMV) isolate from China (SPFMV-CH) associated with one or two other fragments (1.30 and 1.45 kb), suggesting the presence oftwo other potyviruses in these sweetpotato clones. The 1.30- and 1.45- kb amplified fragments were cloned into the Bluescript plasmid and partially sequenced. Comparison of the deduced partial amino acid sequences derived from the amplified fragments with those of the C- terminal region of the nuclear inclusion b protein and the N-terminal region of the coat protein of SPFMV-CH and of several other potyviruses indicated mixed infections by distinct potyviruses in the sweetpotato clones we investigated. Dot blot assays and preliminary sequence analysis indicate that the virus corresponding to the 1.30-kb fragment is closely relate

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