Prunus necrotic ringspot virus (PNRSV) and cherry virus A (CVA) are two viruses that mainly infect plants of the genus Prunus. Full-length sequences of these two viruses, collected in the Czech Republic from Prunus cerasifera plants, were obtained via HTS sequencing. Phylogenetic analyses based on the NJ method and Splitstree tools showed that the Czech PNRSV isolate (ON088600-ON088602) is a divergent isolate from other molecular groups, sharing less than 97% pairwise nucleotide identity with members of other groups. The Czech CVA isolate (ON088603) belonged to molecular subgroup III-2, clustered with isolates from non-cherry hosts, and shared the highest pairwise nucleotide identity (99.7%) with an isolate of Australian origin.

• Klíčová slova
• CVA, PNRSV, biological assay, phylogenetic analysis, symptoms,
• MeSH
• Flexiviridae MeSH
• fylogeneze MeSH
• Ilarvirus MeSH
• nukleotidy MeSH
• Terminalia * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Austrálie MeSH
• Názvy látek
• nukleotidy MeSH

Apple hammerhead viroid (AHVd) was detected in the apple cultivar 'Šampion' and in mixed infection with Solanum nigrum ilarvirus 1 (SnIV-1) in the cultivars 'Selena' and 'Jonagored Supra', using a high-throughput sequencing method. Experiments were conducted to eliminate both pathogens in apples using meristem tip cultures in combination with the antivirotics ribavirin, rimantadine, and zidovudine. Elimination of both pathogens was verified by repeated RT-PCR and qRT-PCR assays after 7-11 months. Elimination of SnIV-1 from all cultivars was successful with each of the three antivirotics at concentrations of 20, 40, and 80 mg L-1. Elimination of AHVd was also achieved, although less effectively and only with ribavirin in the concentration range of 20-160 mg L-1.

• Klíčová slova
• HTS, apple, chemotherapy, viroid, virus,
• MeSH
• antivirové látky farmakologie   MeSH
• Ilarvirus * MeSH
• Malus * MeSH
• ribavirin farmakologie   MeSH
• rimantadin MeSH
• Solanum nigrum * MeSH
• viroidy * MeSH
• zidovudin MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antivirové látky MeSH
• ribavirin MeSH
• rimantadin MeSH
• zidovudin MeSH

Apple mosaic virus (ApMV) is a widespread ssRNA virus which infects diverse species of Rosales. The phylogenetic analysis of complete capsid protein gene of the largest set of ApMV isolates discriminated two main clusters of isolates: one cluster correlates with Maloideae hosts and Trebouxia lichen algae hosts; a second with hop, Prunus, and other woody tree hosts. No correlation was found between clusters and geographic origin of virus isolates, and positive selection hypothesis in distinct hosts was not confirmed: in all virus populations, purifying selection had occurred. GGT→AAT substitution resulted in Gly→Asn change inside the zinc-finger motif in the capsid protein was revealed specific for discrimination of the clusters and we hypothesise that could influence the host preference.

• MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• hostitelská specificita * MeSH
• Ilarvirus klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• Malus virologie   MeSH
• molekulární sekvence - údaje MeSH
• nemoci rostlin virologie   MeSH
• sekvence aminokyselin MeSH
• virové plášťové proteiny genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• virové plášťové proteiny MeSH

A quantitative PCR, real-time RT-PCR, and one-step real-time RT-PCR using SYBR Green-based tools for reliable detection and relative quantitation of Prune dwarf virus (PDV) in stone fruits are described. The assay reliability was tested on 55 samples from different hosts and regions. The sensitivity of the assay was also compared with other assays with different primers. Two plant-expressed genes, actin and 18S rRNA, were used as housekeeping genes for accurate quantitation of PDV in stone fruit trees. The expression of the gene for actin and the 18S ribosomal RNA gene corresponded with each other accurately, with standard deviation values of 1.905 cycles on average, 1.36 for Prunus persica, and 2.45 for other Prunus species tested. The results of this study support the need to use more than one housekeeping gene as an internal control to avoid possible errors caused by unstable internal control gene mRNA expression when quantifying the extent of PDV infection.

• MeSH
• aktiny genetika   MeSH
• barvení a značení metody   MeSH
• benzothiazoly MeSH
• chinoliny MeSH
• diaminy MeSH
• fluorescenční barviva MeSH
• Ilarvirus genetika   izolace a purifikace   MeSH
• organické látky MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   normy   MeSH
• referenční standardy MeSH
• RNA ribozomální 18S genetika   MeSH
• RNA rostlin genetika   MeSH
• RNA virová genetika   MeSH
• rostlinné proteiny genetika   MeSH
• slivoň virologie   MeSH
• virová nálož * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• benzothiazoly MeSH
• chinoliny MeSH
• diaminy MeSH
• fluorescenční barviva MeSH
• organické látky MeSH
• RNA ribozomální 18S MeSH
• RNA rostlin MeSH
• RNA virová MeSH
• rostlinné proteiny MeSH
• SYBR Green I MeSH Prohlížeč

Eight new sequences of European isolates from almond, apple, hop, prune and pear of the Apple mosaic ilarvirus (ApMV) capsid protein gene are presented. A consensus sequence was established as having 654 nucleotides (nt) and two American and two European isolates were identified to have insertions 6 to 15 nucleotides after nt position 141. The insertion resulted in the American isolate A inframeshift repaired with two point insertions 17 and 68 nt downstream. The RNA around the insertion point can potentially form a stable secondary structure with three hairpins. The insertions could stabilise this structure or could be neutral. The predicted folding of the translated protein is not influenced by the insertions or frameshift, and we speculate that the region after nt position 141 is without reasonable selection pressure and represents a hot spot for the accumulation of insertion mutations in ApMV.

• MeSH
• genetická variace MeSH
• Humulus virologie   MeSH
• Ilarvirus chemie   genetika   MeSH
• inzerční mutageneze MeSH
• kapsida chemie   genetika   MeSH
• konformace nukleové kyseliny MeSH
• Malus virologie   MeSH
• molekulární sekvence - údaje MeSH
• RNA virová chemie   MeSH
• sekundární struktura proteinů MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční seřazení MeSH
• slivoň virologie   MeSH
• virové geny * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• RNA virová MeSH

The coat protein (CP) gene of Prunus necrotic ringspot virus (PNRSV) was cloned into pET 16b vector and expressed in Escherichia coli. CP-enriched fractions were prepared from whole cell lysate by differential centrifugation. The fraction sedimenting at 20,000 x g for 30 mins was used for preparation of a rabbit antiserum to CP. This antiserum had a titer of 1:2048 and reacted in a double-antibody sandwich ELISA (DAS-ELISA).

• MeSH
• Ilarvirus genetika   izolace a purifikace   MeSH
• kapsida genetika   izolace a purifikace   MeSH
• klonování DNA MeSH
• molekulární sekvence - údaje MeSH
• rekombinantní proteiny genetika   izolace a purifikace   MeSH
• sekvence nukleotidů MeSH
• stromy virologie   MeSH
• virové geny MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• rekombinantní proteiny MeSH

The 3'-part of the movement protein gene, the intergenic region and the complete coat protein gene of sixteen isolates of Prunus necrotic ringspot virus (PNRSV) from five different host species from the Czech Republic were sequenced in order to search for the bases of extensive variability of viroses caused by this pathogen. According to phylogenetic analyses all the 46 isolates sequenced to date split into three main groups, which correlated to a certain extend with their geographic origin. Modelled serological properties showed that all the new isolates belong to one serotype.

• MeSH
• fylogeneze MeSH
• Ilarvirus klasifikace   genetika   MeSH
• kapsida analýza   chemie   MeSH
• molekulární sekvence - údaje MeSH
• otevřené čtecí rámce MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• virové proteiny genetika   MeSH
• viry rostlin - pohybové proteiny MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• virové proteiny MeSH
• viry rostlin - pohybové proteiny MeSH

Thirteen cultivars of hop (Humulus lupulus L.) were tested by reverse transcription-polymerase chain reaction (RT-PCR) for the presence of apple mosaic virus (ApMV). The virus was detected in various amounts in all tested cultivars. Control hop clones derived from tissue cultures, treated by thermotherapy and maintained in greenhouse were virus-free. The procedure for sample preparation and RT-PCR of ApMV is described in detail.

• MeSH
• Ilarvirus genetika   MeSH
• molekulární sekvence - údaje MeSH
• nemoci rostlin virologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• RNA virová analýza   MeSH
• rostliny virologie   MeSH
• sekvence nukleotidů MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• RNA virová MeSH