The wheat streak mosaic virus (WSMV), vectored by the wheat curl mite, is globally distributed and threatens wheat production worldwide. Since its first occurrence in Serbia in the 1960s, WSMV presence has not been monitored. In 2019, a total of 62 samples of fi ve wheat cultivars from eight locations in Serbia were collected and tested for the presence of nine common wheat viruses: WSMV, barley yellow dwarf virus-PAV, -MAV, -SGV, and -RMV, cereal yellow dwarf virus-RPV, wheat spindle streak virus, brome mosaic virus, and soil-borne wheat mosaic virus, using individual or multiplex RT-PCR. WSMV was detected in 58.1% of the tested samples in seven wheat crops at five different locations. Species-specific primers failed to detect the presence of the other eight tested viruses. For further confirmation of WSMV, RT-PCR with the WS8166F/WS8909R primers covering the coat protein (CP) gene was carried out for both amplification and sequencing. The amplified product of the correct predicted size (750 bp) derived from four selected isolates, 98-19, 99-19, 102-19 and 120-19, was sequenced and deposited in GenBank (MT461299, MT461300, MT461301 and MT461302, respectively). Serbian WSMV isolates showed very high nucleotide identity (98.16-99.02%) and shared a deletion of triplet codon GCA at nucleotide position 8412- 8414 resulting in deletion of glycine amino acid (Gly2761). Phylogenetic analysis conducted on CP gene sequences revealed the existence of four clades, named A, B, C and D, and one recently introduced clade B1. All Serbian wheat WSMV isolates grouped into clade B together with other European isolates and one isolate from Iran. The results of this study provide the first insight into molecular characterisation of Serbian WSMV isolates, indicating their close relationship with other European isolates and existence of a single genotype in the country. Phylogenetic analysis also confirms the dispersal of WSMV isolates throughout Europe from a single locus.
The present study provides a new insight into the existing Xanthomonas arboricola pv. pruni (Xap) bacterial population originating from peach and apricot in two Western Balkans countries (Serbia and Montenegro). Multilocus sequence typing and analysis of the sequences of nine housekeeping genes revealed homology between the tested Xap strains as well as with the European population of this bacterium. The tested strains share the same haplotype (Haplotype I) with the Xap strains from Italy, France, Spain, United States, Australia and Brazil. The revealed single nucleotide change (G ↔ C) in the sequences of the gyrB1 gene differentiates Haplotype I from Haplotype II (Xap from South Korea, New Zealand, Argentina and Uruguay). The detached‐leaf bioassay results confirmed differences in virulence between strains originating from peach and apricot towards Prunus armeniaca (apricot), indicating host specialization of the apricot strain towards this host. For the first time, immunity of P. fruticosa (European ground cherry) to Xap was established. According to the AUDPC, PCA and cluster analysis, other Prunus spp. were classified as having low susceptibility (P. mahaleb, P. cerasus and P. avium), as susceptible (P. domestica) and as highly susceptible (P. persica, P. dulcis, P. cerasifera and P. spinosa). Xap strains were also found to be susceptible to 10 tested antibiotics. This study provides valuable knowledge on the Xap population from stone fruit grown in the Western Balkans region as well as the source of immunity, which could serve as a starting point for breeding Prunus cultivars and could be used as the main control strategy.
The fauna of aphids appearing on cucurbits, including oilseed pumpkin have been poorly investigated in Serbia. Yellow water traps were used to determine the diversity of aphid species visiting cucurbits in Serbia and to monitor their flight activity. During the years 2009 to 2011, a total of 1,447 specimens were collected and a total of 57 different aphid taxa were identified. In 2009, the highest total number of aphids were caught (755), followed by 203 and 489 aphids in 2010 and 2011, respectively. The most abundant species were Aphis fabae (15%), Myizus persicae (13.4%) and Acyrthosiphon pisum (11.7%), followed by Anoecia corni (6.4%) and Aphis gossypii (5.6%). All of them, except A. corni, are reported as efficient vectors of cucurbit viruses. These five species represent 52.1% of all aphid species collected within this study. Out of 57 taxa found in oilseed pumpkin crops, 17 are known as vectors of ZYMV, 18 of WMV and 15 of CMV. Generally, 25 out of a total of 57 taxa are known as vectors of at least one of three viruses present in Serbia. From total of 1,447 specimens collected during three years of investigation, 917 (63.4%) are vectors of at least one of three viruses. Individual oilseed pumpkin plants were frequently colonized by A. gossypii during all three years of investigation. Results of this study showed that Serbian oilseed pumpkin growing localities are grouped among medium to reach in aphidofauna, according to total Shannon-Weaver index values which varied from 1.8 to 2.8. The highest value of Shannon-Weaver (2.8) was recorded in the Bački Petrovac I locality in 2009, while the lowest Shannon-Weaver (1.8) was recorded in Kisač locality in 2010. The maximum values of Shanon-Weaver diversity index in almost all crops were recorded in the second week of investigation or in early stages of oilseed pumpkin growth when plants had 5-6 true leaves or were in flowering stage, when potential virus infection could lead to significant yield losses.
Aim of study: To report the occurrence of Pepino mosaic virus (PepMV) on tomato in Serbia and to genetically characterize Serbian PepMV isolates.Area of study: Tomato samples showing virus-like symptoms were collected in the Bogojevce locality (Jablanica District, Serbia).Material and methods: Collected tomato samples were assayed by DAS-ELISA using antisera against eight economically important or quarantine tomato viruses. Three selected isolates of naturally infected tomato plants were mechanically transmitted to tomato ‘Novosadski jabučar’ seedlings. For confirmation of PepMV infection, RT-PCR was performed using specific primers PepMV TGB F/PepMV UTR R. Maximum-likelihood phylogenetic tree was constructed with 47 complete CP gene sequences of PepMV to determine the genetic relationship of Serbian PepMV isolates with those from other parts of the world.Main results: The results of DAS-ELISA indicated the presence of PepMV in all tested samples. Mechanically inoculated ‘Novosadski jabučar’ seedlings expressed yellow spots and light and dark green patches, bubbling, and curled leaves. All tested tomato plants were RT-PCR positive for the presence of PepMV. The CP sequence analysis revealed that the Serbian PepMV isolates were completely identical among themselves and shared the highest nucleotide identity of 95.1% (99.2% aa identity) with isolate from Spain (FJ263341). Phylogenetic analysis showed clustering of the Serbian PepMV isolates into CH2 strain, but they formed separate subgroup within CH2 strain.Research highlights: This is the first data of the presence of PepMV in protected tomato production in Serbia. Considering increased incidence and rapid spread in Europe, the presence of PepMV on tomato could therefore represent serious threat to this valuable crop in Serbia.
Grapevine rupestris stem pitting-associated virus (GRSPaV), a member of the genus Foveavirus, is a commonly detected grapevine virus around the world. Despite the broad and global distribution of GRSPaV, no information on its presence in Serbia had been available before this study. In October 2018, 3-year-old grapevine plants of 'Cabernet Sauvignon' showing reduced growth and leaf reddening were observed in the vicinity of Krnjevo locality. The results of an mRT-PCR test showed that only samples prepared from cortical scrapings of mature symptomatic grapevine canes yielded an amplicon of 155 bp that confirmed the presence of GRSPaV. No amplification products were recorded in samples prepared from basal leaves or petioles. A serological assay confirmed the presence of GRSPaV in all tested mature cane samples. For further analyses, the CP (coat protein) genes of two selected isolates (VL8 and VL15) of naturally infected grapevine canes were partially sequenced and six sequence variants were determined, showing a certain variability among them. The constructed phylogenetic tree revealed the grouping of Serbian sequence variants into separate subgroups within the same phylogenetic lineage. This is the first report of GRSPaV present in Serbia. Further monitoring and increased phytosanitary control measures are needed to prevent potential spreading and introduction of new virus variants.
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