Frequency and Molecular Characterization of Watermelon Mosaic Virus from Serbia
Učestalost pojave i molekularna karakterizacija virusa mozaika lubenice poreklom iz Srbije
View/ Open
Author
Vucurovic, Ana
Bulajic, Aleksandra
Stankovic, Ivana
Ristic, Danijela
Keywords
PumpkinWatermelon mosaic virus
Molecular identification
Genetic diversity
Phylogeny
Tikve
virus mozaika lubenice
molekularna identifikacija
genetièki diverzitet
filogenetska analiza
Metadata
Show full item recordAbstract
Watermelon mosaic virus (WMV) is widespread in cucurbit crops, most commonly occuring
in temperate and Mediterranean regions. In Serbia WMV has been detected in single and
mixed infections with Zucchini yellow mosaic virus and Cucumber mosaic virus in field-grown
pumpkin and squash crops. Among pumpkin-affecting viruses WMV is the most frequent
one, both by the number of localities and its incidence at each location. During the growing
season of 2009, samples from 583 plants of Cucurbita pepo cvs. Olinka, Belgrade zucchini
and Tosca (Zucchini group), as well as from C. maxima and C. moschata showing symptoms
of virus infection were collected from 12 commercial fields at eight localities and analyzed
by DAS-ELISA using polyclonal antisera specific to six most important cucurbit viruses.
Interestingly, WMV was detected at fewer sites and had lower incidence rate than in two previous
years. In single infections, WMV was found in 11% of tested plants in three fields; in mixed
infections with ZYMV, it was recorded in 9.9% of plants in five fields and with CMV in only 0.2%
in one field. The partial coat protein gene and 3’ non-translated region from two representative
isolates of WMV originating from different localities and host plant species were amplified
by RT-PCR, sequenced, and compared with the sequences available in GenBank database.
The PCR-amplified fragment of predicted size of approximately 1017 bp was obtained. The
sequences of isolates 137-08 (Acc. No. GQ259958) and 159-08 (GU144020) proved to be 94-99%
identical at the nucleotide level with those from other parts of the world. The sequences of
these two isolates differed from each other only at two nucleotide positions, without any amino
acid substitution. Phylogenetic analysis of 57 isolates based on 750 bp sequences of the
coat protein gene showed no correlation between isolates and their geographic origin, and it
also indicated that these isolates fell into three molecular groups of strains. Serbian WMV isolates
belong to group I, the most widespread group of isolates of this virus in the world. Virus mozaika lubenice (Watermelon mosaic virus, WMV) široko je rasprostranjen u usevima
vrežastih kultura, mada se najčešće javlja na Mediteranu i regionima sa umerenom klimom.
U Srbiji, WMV je detektovan u pojedinačnim i mešanim infekcijama sa virusom žutog mozaika
cukinija (Zucchini yellow mosaic virus, ZYMV) i virusom mozaika krastavca (Cucumber mosaic
virus, CMV) u usevima tikava i bundeva gajenim na otvorenom polju. Od virusa tikava, WMV
je često prevalentan virus, kako po broju lokaliteta u kojima je dokazan, tako i po učestalosti
u samom usevu. Iz ukupno 12 komercijalnih useva na osam lokaliteta, 2009. godine sakupljeno
je 583 biljaka Cucurbita pepo cvs. Olinka, Beogradska tikvica i Tosca (cukini forma), kao i C.
maxima i C. moschata, sa simptomima virusnih zaraza, koje su analizirane na prisustvo šest najznačajnijih
virusa tikava primenom DAS-ELISA metode. WMV je detektovan na manjem broju
lokaliteta i u manjoj učestalosti u poređenju sa prethodne dve vegetacije. U pojedinačnim
infekcijama, WMV je detektovan u 11% testiranih biljaka u tri useva, a u mešanim sa ZYMV u
9,9% testiranih biljaka u pet useva i sa CMV u 0,2% testiranih biljaka u jednom usevu. Deo gena
za protein omotača virusa i deo 3’ neprepisujućeg regiona dva odabrana izolata WMV poreklom
iz različitih lokaliteta i biljaka domaćina umnožen je primenom RT-PCR metode, sekvencioniran
i upoređen sa sekvencama dostupnim u GenBank bazi podataka. Dobijen je PCR
produkt očekivane veličine od 1017 bp. Sekvence izolata 137-08 (Acc. No. GQ259958) i 159-08
(GU144020) pokazale su 94-99% nukleotidne identičnosti sa izolatima WMV iz drugih delova
sveta. Sekvence ova dva izolata razlikovala su se jedna od druge samo u dva nukleotidna mesta
što nije imalo za posledicu aminokiselinske supstitucije. Filogenetske analize 57 izolata, na
osnovu 750 bp sekvence gena za protein omotača, nisu ukazale na korelaciju sa geografskim
poreklom izolata, a pokazala su grupisanje izolata u tri molekularne grupe sojeva. Izolati WMV
iz Srbije pripadaju grupi I, grupi koja obuhvata najrasprostranjenije izolate ovog virusa u svetu.