Presence and molecular characterization of cucumber mosaic virus on safflower in Serbia

Abstract

Safflower (Carthamus tinctorius L.) is an important oilseed crop belonging to the family Asteraceae. A total of 46 safflower samples were collected from Srbobran locality (South Bačka District) in Serbia in 2015 and analysed for the presence of cucumber mosaic virus (CMV), alfalfa mosaic virus (AMV), and lettuce mosaic virus (LMV), using commercial double-antibody sandwich (DAS)-ELISA kits. Both viruses, CMV and AMV, were detected serologically in the collected samples. None of the analysed samples was found to be positive for LMV. The presence of CMV was further confirmed by mechanical transmission to test the plants including Chenopodium quinoa, C. amaranticolor, Nicotiana glutinosa, and Datura stramonium as well as C. tinctorius, confirming the infectious nature of the disease. Molecular detection of CMV was performed by amplification of a 871 bp fragment in all the tested samples, using the specific primers CMVCPfwd/CMVCPrev that amplify the entire coat protein (CP) gene and part of 3'and 5'-UTRs of CMV RNA 3. The RT-PCR products derived from the isolates 290Saff and 294Saff were sequenced (MH577791 and MH577792, respectively) and compared with the CMV sequences available in GenBank. Phylogenetic analysis based on CP gene sequences showed clustering of the selected isolates into three subgroups: IA, IB and II. Serbian CMV isolates found in safflower belong to subgroup II. To our knowledge, this is the first report on CMV infection of safflower in Serbia, which has the potential to cause substantial damage to safflower production and pose a threat to other economic crops grown in Serbia.

Šafranika (Carthamus tinctorius L.) predstavlja jednu od važnih uljanih kultura koja pripada familiji Asteraceae. Tokom 2015. godine, prikupljeno je 46 uzoraka biljaka šafranike poreklom iz Srbobrana koji su serološki testirani na prisustvo virusa mozaika krastavca (cucumber mosaic virus, CMV), virusa mozaika lucerke (alfalfa mosaic virus, AMV) i virusa mozaika salate (lettuce mosaic virus, LMV) korišćenjem komercijalnih kitova za DAS-ELISA test. U prikupljenim uzorcima dokazano je prisustvo CMV i AMV, dok prisustvo LMV nije dokazano ni u jednom od testiranih uzoraka. Prisustvo CMV je dalje potvrđeno mehaničkim inokulacijama test biljaka Chenopodium quinoa, C. amaranticolor, Nicotianaglutinosa i Datura stramonium kao i na C. tinctorius, čime je potvrđena infektivna priroda oboljenja. Molekularna detekcija CMV obavljena je amplifikacijom fragmenta dužine 871 bp kod svih ispitivanih izolata korišćenjem specifičnih prajmera CMVCPfwd/CMVCPrev koji omogućavaju umnožavanje celog CP gena kao i delove 5' i 3' UTR. U cilju dalje identifikacije, RT-PCR produkti izolata 290Saff i 294Saff su sekvencionisani (MH577791 i MH577792) i upoređeni sa CMV sekvencama dostupnim u GenBank bazi podataka. Filogenetska analiza na osnovu sekvence CP gena pokazala je grupisanje odabranih izolata u tri podgrupe, IA, IB i II. Izolati CMV iz šafranike poreklom iz Srbije grupisali su se u podgrupu II. Prema našim saznanjima, ovo je prvi izveštaj o prisustvu CMV na biljkama šafranike u Srbiji koji može da nanese veliku štetu u proizvodnji ove biljne vrste, a takođe predstavlja pretnju drugim ekonomski značajnim gajenim biljnim vrstama u Srbiji.