Antiviral activity of acyclic nucleoside phosphonates PMEA, (S)-HPMPC, PMEDAP and ribavirin against Cauliflower mosaic virus in Brassica pekinensis

  • Josef Špak
  • Ivan Votruba
  • Daniela Pavingerová
  • Antonín Holý
  • Vlastimila Špaková
  • Karel Petrzik
Original Paper


Antiviral effects of acyclic nucleoside phosphonates PMEA, (S)-HPMPC, PMEDAP, and ribavirin on double-stranded DNA Cauliflower mosaic virus (CaMV) were evaluated in Brassica pekinensis plants grown in vitro on liquid medium. A double-antibody sandwich ELISA was used for relative quantification of viral protein and PCR for detection of CaMV nucleic acid in plants. Ribavirin and PMEA had no significant antiviral effect. (S)-HPMPC at concentration 50 mg l−1 and PMEDAP at concentrations 50 and 12.5 mg l−1 significantly (P < 0.05) reduced CaMV concentration in plants within 42–63 days to levels detectable neither by ELISA nor by PCR. A phytotoxicity experiment resulted in progressive yellowing of leaves and dwarfing in plants cultured 42 days on media with concentrations 12.5, 25 and 50 mg l−1 of (S)-HPMPC and PMEDAP. Reduction in fresh and dry weights of plants was significant (P < 0.05) already at 12.5 mg l−1 with both compounds.


Caulimovirus dsDNA Chemotherapy Pararetrovirus Phytotoxicity Virus eradication 



Acyclic nucleoside phosphonate


Cauliflower mosaic virus


Murashige and Skoog medium











This project was funded by grants nos. 522/09/0707 from the Grant Agency of the Czech Republic and AV0Z50510513 from the Academy of Sciences of the Czech Republic, as well as by Research Project OZ40550506 of the Institute of Organic Chemistry and Biochemistry. The authors thank Dr. I. Dostálková, Ph.D., Faculty of Science, University of South Bohemia, for consultations on statistical processing of the data.


  1. Acquadro A, Papanice MA, Lanteri S, Bottalico G, Portis E, Campanale A, Finetti-Sialer MM, Mascia T, Sumerano P, Gallitelli D (2010) Production and fingerprinting of virus-free clones in a reflowering globe artichoke. Plant Cell Tissue Organ Cult 100:329–337CrossRefGoogle Scholar
  2. Cheong EJ, Mock R, Li R (2012) Elimination of five viruses from sugarcane using in vitro culture of axillary buds and apical meristems. Plant Cell Tissue Organ Cult. doi: 10.1007/s11240-011-0108-3 Google Scholar
  3. De Clercq E (2011) The clinical potential of the acyclic (and cyclic) nucleotide phosphonates. The magic of the phosphonate bond. Biochem Pharmacol 82:99–109PubMedCrossRefGoogle Scholar
  4. Hansen AJ (1989) Antiviral chemicals for plant disease control. Crit Rev Plant Sci 8:45–88CrossRefGoogle Scholar
  5. Hanzliková-Vašková D, Špak J, Petrzik K (2006) Variability in sequence of Strawberry vein banding virus. Biol Plant 50:660–666CrossRefGoogle Scholar
  6. Helliot B, Panis B, Frison E, De Clercq E, Swennen R, Lepoivre P, Neyts J (2003) The acyclic nucleoside phosphonate analogues, adefovir, tenofovir and PMEDAP, efficiently eliminate banana streak virus from banana (Musa ssp.). Antiviral Res 59:121–126PubMedCrossRefGoogle Scholar
  7. Holý A, Rosenberg I, Dvořáková H (1989) Synthesis of N-(2-phosphonylmethoxy-ethyl) derivatives of heterocyclic bases. Collect Czech Chem Commun 54:2190–2210CrossRefGoogle Scholar
  8. Murashige TF, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  9. Ramgareeb S, Snyman SJ, van Antwerpen T, Rutherford RS (2010) Elimination of virus and rapid propagation of disease-free sugarcane (Saccharum spp. cultivar NCo376) using apical meristem culture. Plant Cell Tissue Organ Cult 100:175–181CrossRefGoogle Scholar
  10. Rommens CMT, Kneppers TJA, Haring MA, Nijkamp HJJ, Hille J (1991) A transposon tagging strategy with/Ac/on plant cell level in heterologous plant species. Plant Sci 74:99–106CrossRefGoogle Scholar
  11. Špak J (1989a) Characterisation of Czechoslovak isolates of Cauliflower mosaic virus from brassica crops. Ochrana rostlin 25:177–181Google Scholar
  12. Špak J (1989b) Purification and serodiagnosis of Cauliflower mosaic virus. Ochrana rostlin 25:241–247Google Scholar
  13. Špak J, Holý A, Pavingerová D, Votruba I, Špaková V, Petrzik K (2010) New in vitro method for evaluating antiviral activity of acyclic nucleoside phosphonates against plant viruses. Antiviral Res 88:296–303PubMedCrossRefGoogle Scholar
  14. Špak J, Votruba I, Pavingerová D, Holý A, Špaková V, Petrzik K (2011) Antiviral activity of tenofovir against Cauliflower mosaic virus and its metabolism in B. pekinensis plants. Antiviral Res 92:378–381PubMedCrossRefGoogle Scholar
  15. Tan R, Wang L, Hong N, Wang G (2010) Enhanced efficiency of virus eradication following thermotherapy of shoot-tip cultures of pear. Plant Cell Tissue Organ Cult 101:229–235CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Josef Špak
    • 1
  • Ivan Votruba
    • 2
  • Daniela Pavingerová
    • 1
  • Antonín Holý
    • 2
  • Vlastimila Špaková
    • 1
  • Karel Petrzik
    • 1
  1. 1.Institute of Plant Molecular BiologyBiology Centre of Academy of Sciences of the Czech Republic, v.v.i.České BudějoviceCzech Republic
  2. 2.Institute of Organic Chemistry and BiochemistryAcademy of Sciences of the Czech Republic, v.v.i.PragueCzech Republic

Personalised recommendations