Detection and partial molecular characterization of grapevine virus A form herbaceous hosts

Vinova loza je domaćin velikog broja virusa od kojih je jedan od ekonomski najznačajnijih A-virus vinove loze (Grapevine virus A, GVA), a osim sadnim materijalom može se širiti štitastim ušima na vinovu lozu i na zeljaste domaćine. Za potrebe ovog rada istraženi su prijenos i prisutnost GVA u zeljastim biljkama metodama ELISA, konvencionalnim RT-PCR-om i PCR-om u realnom vremenu. Pozitivne zeljaste biljke na GVA bile su oštrodlakavi šćir (Amaranthus retroflexus), Nicotiana benthamiana, loboda kamenjarka (Chenopodium murale) i mak turčinjak (Papaver rhoeas). Odabrano je 9 GVA izolata koji su sekvencirani, a dobivene sekvence uspoređene su međusobno te s drugim izolatima iz GenBanke. Dobivene sekvence razlikuju se u nukleotidnom slijedu gdje se sastoje od 327 monomorfnih i 63 polimorfnih mjesta, dok na aminokiselinskoj razini nije došlo do značajnih promjena. Sekvence izvorišne biljke dokazale su prisutnost više GVA varijanata. Na filogenetskom stablu, istraživani GVA izolati grupirali su se u dvije zasebne skupine.The grapevine can be attacked by a large number of viruses of which the most important is Grapevine virus A (GVA) which can be spread with both planting materials and by mealybugs on grapes and herbaceous hosts. For the purposes of this research, the transmission and presence of GVA in herbaceous plants were investigated by ELISA, conventional RT-PCR and Real-Time PCR methods. Plants that tested positive for GVA were redroot pigweed (Amaranthus retroflexus), Nicotiana benthamiana, nettle-leaved goosefoot (Chenopodium murale) and common poppy (Papaver rhoeas). Nine isolates were selected and sent for sequencing. The obtained sequences were compared with each other and with other isolates from GenBank. The resulting sequences differ in the nucleotide sequence; they consist of 327 monomorphic and 63 polymorphic sites, while at the amino acid level there were no significant changes. Source plant sequences proved the presence of multiple GVA variants. Within the phylogenetic tree tested GVA isolates were gathered in two distinct groups