The antiviral effect of the acyclic nucleoside phosphonate tenofovir (R)-PMPA on double-stranded DNA Cauliflower mosaic virus (CaMV) in Brassica pekinensis plants grown in vitro on liquid medium was evaluated. Double antibody sandwich ELISA and PCR were used for relative quantification of viral protein and detecting nucleic acid in plants. (R)-PMPA at concentrations of 25 and 50 mg/l significantly reduced CaMV titers in plants within 6-9 weeks to levels detectable neither by ELISA nor by PCR. Virus-free plants were obtained after 3-month cultivation of meristem tips on semisolid medium containing 50 mg/l (R)-PMPA and their regeneration to whole plants in the greenhouse. Studying the metabolism of (R)-PMPA in B. pekinensis revealed that mono- and diphosphate, structural analogs of NDP and/or NTP, are the only metabolites formed. The data indicate very low substrate activity of the enzymes toward (R)-PMPA as substrate. The extent of phosphorylation in the plant's leaves represents only 4.5% of applied labeled (R)-PMPA. In roots, we detected no radioactive peaks of phosphorylated metabolites of (R)-PMPAp or (R)-PMPApp.

• MeSH
• adenin analogy a deriváty   metabolismus   farmakologie   MeSH
• antivirové látky metabolismus   farmakologie   MeSH
• biotransformace MeSH
• Brassica metabolismus   virologie   MeSH
• Caulimovirus účinky léků   růst a vývoj   MeSH
• DNA virů analýza   MeSH
• ELISA metody   MeSH
• kyseliny fosforité metabolismus   farmakologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• virová nálož MeSH
• virové proteiny analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

A new method was developed for testing antiviral compounds against plant viruses based on rapidly growing brassicas in vitro on liquid medium. This method enables exchange of media containing tested chemicals in various concentrations and simultaneous evaluation of their phytotoxicity and antiviral activity. While using ribavirin as a standard for comparison, phytotoxicity and ability of the acyclic nucleotide analogues (R)-PMPA, PMEA, PMEDAP, and (S)-HPMPC to eliminate ssRNA Turnip yellow mosaic virus (TYMV) were evaluated by this method. Double antibody sandwich ELISA and real-time PCR were used for relative quantification of viral protein and nucleic acid in plants. Ribavirin had the most powerful antiviral effect against TYMV. On the other hand, (R)-PMPA and PMEA had no antiviral effect and almost no phytotoxicity compared to the control. (S)-HPMPC and PMEDAP showed moderate antiviral effect, accompanied by higher phytotoxicity. The tested compounds can be screened within 6-9 weeks in contrast to the 6 months for traditionally used explants on solid medium. The method enables large-scale screening of potential antivirals for in vitro elimination of viruses from vegetatively propagated crops and ornamentals.

• MeSH
• acyklické uhlovodíky farmakologie   terapeutické užití   MeSH
• antivirové látky farmakologie   terapeutické užití   MeSH
• Brassica účinky léků   růst a vývoj   virologie   MeSH
• hydroponie metody   MeSH
• kultivační média MeSH
• mikrobiální testy citlivosti metody   MeSH
• nemoci rostlin terapie   virologie   MeSH
• nukleosidy farmakologie   terapeutické užití   MeSH
• replikace viru účinky léků   MeSH
• ribavirin farmakologie   terapeutické užití   MeSH
• Tymovirus účinky léků   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH