Advertisement

European Journal of Plant Pathology

, Volume 108, Issue 4, pp 367–371 | Cite as

Development of a General Potexvirus Detection Method

  • René A.A. van der Vlugt
  • Miranda Berendsen
Article

Abstract

A method was developed for the detection of viruses from the genus Potexvirus. Following alignment of full-length RNA sequences and deduced amino acid sequences of 10 different viruses from the genus Potexvirus, a number of conserved sequence motifs were identified in the viral replicase-encoding region. Seven different primers based on these motifs were tested for their efficiency as potexvirus group-specific cDNA and/or PCR primer. Several combinations of primers proved capable of generating DNA fragments for each of six different potexviruses tested. One cDNA primer in combination with one PCR primers set proved most successful in reliably generating discrete PCR products. Application of this set to a number of different potexviruses, some for which no sequence data had been published yet, resulted in amplification of virus-specific PCR products for all viruses tested. Sequence analysis of cloned PCR products confirmed their identity. This general potexvirus primer set can be useful for the identification of (unknown) potexvirus infections.

PCR-detection polymerase potexvirus virus identification 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Brunt AA, Crabtree K, Dallwitz MJ, Gibbs AJ, Watson L and Zurcher EJ (eds) (1996) PlantViruses Online: Descriptions and Lists from the VIDE Database. Version: 16th January 1997Google Scholar
  2. Chen J, Chen J and Adams MJ (2001) A universal PCR primer to detect members of the Potyviridae and its use to examine the taxonomic status of several members of the family. Archives of Virology 146: 757–766Google Scholar
  3. Gibbs A and Mackenzie A (1997) A primer pair for amplyfing part of the genome of potyvirids by RT-PCRT. Journal ofVirological methods 63: 9–16Google Scholar
  4. Jones RAC, Koenig R and Lesemann D-E (1980) Pepino mosaic virus, a new potexvirus from pepino (Solanum muricatum). Annals of Applied Biology 94: 61–68Google Scholar
  5. Langeveld SA, Dore J-M, Memmelink J, Derks AFLM, Van der Vlugt CIM, Asjes CJ and Bol JF (1991) Identification of potyviruses using the polymerase chain reaction with degenerate primers. Journal of General Virology 76: 1531–1541Google Scholar
  6. Liu H-J, Giambrone JJ and Dormitorio T 1994 Detection of genetic variations in serotype I isolates of infectious bursal disease virus using polymerase chain reaction and restriction endonuclease analysis. Journal of Virological Methods 48: 281–291Google Scholar
  7. Mentaberry A and Orman B (1995) Potexviruses. In: Sing RP, Singh US and Kohmoto K (eds) Pathogenesis and Host Specificity in Plant Diseases. Histopathological, Biochemical, Genetic and Molecular bases. Vol 3: Viruses and Viroids (pp 19–33). Elsevier Science Ltd, Kidlington, UKGoogle Scholar
  8. Pappu SS, Brand R, Pappu HR, Rybicki EP, Gough KH, Frenkel MJ and Niblett CL (1993) A polymerase chain reaction method adapted for selective amplification and cloning of 3′ sequences of potyviral genomes: application to dasheen mosaic virus. Journal of Virological Methods 41: 9–20Google Scholar
  9. Tian T, Klaasen VA, Soong J, Wisler G, Duffus JE and Falk BW (1996) Generation of cDNAs specific to lettuce infectious yellows closterovirus and other white fly transmitted viruses by RT-PCR and degenerate oligonucleotide primers corresponding to the closterovirus gene encoding the heat shock protein homolog. Phytopathology 86: 1167–1173Google Scholar
  10. van der Vlugt RAA, Steffens P, Cuperus C, Barg E, Lesemann D-E, Bos L and Vetten HJ (1999) Further evidence that shallot yellow stripe virus (SYSV) is a distinct potyvirus and reidentification of Welsh onion yellow stripe virus as a SYSV strain. Phytopathology 89: 148–155Google Scholar
  11. van der Vlugt RAA, Stijger CCMM, Verhoeven JThJ and Lesemann D-E (2000) First Report of Pepino Mosaic Virus on Tomato. Plant Disease 84: 103Google Scholar
  12. van Regenmortel MHV, Fauquet CM, Bishop DHL, Carstens EB, Estes MK, Lemon SM, Maniloff J, Mayo MA, McGeoch DJ, Pringle CR and Wickner RB (eds) (2000) In: Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses (pp 975–981) Academic Press, San DiegoGoogle Scholar
  13. Verwoerd TC, Dekker BMM and Hoekema A (1989) A small scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Research 17: 2362Google Scholar
  14. Vetten HJ, Butgereitt A and Winter S (1997) Differentiation of Allium potyviruses by polymerase chain reaction. In:Dehne H-W, Adam G, Diekmann M, Frahm J, Mauler-Machnik A and van Halteren P (eds) Diagnosis and Identification of Plant Pathogens (pp 469–471) Kluwer Academic Publishers, DordrechtGoogle Scholar
  15. Winter S, Körbler M and Köllner U (1997) Detection and differentiation of sweet potato closteroviruses by RT-PCR and single-strand conformation polymorphism. In: Dehne H-W, Adam G, Diekmann M, Frahm J, Mauler-Machnik A and van Halteren P (eds) Diagnosis and Identification of Plant Pathogens (pp 473–475) Kluwer Academic Publishers, DordrechtGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • René A.A. van der Vlugt
    • 1
  • Miranda Berendsen
    • 1
  1. 1.Plant Research International B.V.WageningenThe Netherlands

Personalised recommendations