Survey of five major grapevine viruses infecting Blatina and Žilavka cultivars in Bosnia and Herzegovina

Abstract

The sanitary status of grapevines has not yet been considered sufficiently in vineyards throughout Bosnia and Herzegovina (BiH). An extensive survey of five major grapevine viruses in the country was carried out in 2019. A total of 630 samples from the two dominant autochthonous cultivars, named Žilavka and Blatina, were tested by DAS-ELISA for the presence of grapevine leafroll-associated viruses (GLRaV-1 and 3), grapevine fleck virus (GFkV), grapevine fanleaf virus (GFLV) and Arabis mosaic virus (ArMV). Eighty-eight % of the samples were positive for at least one virus, and all five viruses were detected, thought with different incidence, i.e. GLRaV-3 (84%), GFLV (43%), GLRaV-1 (14%), GFkV (10%) and ArMV (0.2%). The majority of infected plants (about 75%) were asymptomatic. Specific virus symptoms were observed in the remaining infected plants, together with the reported GLRaV vectors, Planococcus ficus and Parthenolecanium corni, while nematodes of the Xiphinema genus were not found in the GFLV- or ArMV-infected vineyards. The GLRaV-3 CP phylogenetic analyses showed 75–100% nucleotide identity between the BiH and reference isolates, and the BiH isolates clustered into the major group. The dNS/dS ratio indicated a negative selection of the virus population, and the lack of geographical structuring within the population was observed. In addition, putative GLRaV-3 recombinants with breakpoints in the 5’ of the CP gene were detected, while no recombinant strains were identified for the other four viruses. The obtained results indicate a deteriorated sanitary status of the cultivated grapevines, the prevalence and intraspecies genetic diversity of GLRaV-3 throughout the country. The establishment of certified grapevine material and adequate virus vector control is therefore of primary importance to prevent further spread of these viruses. This study presents the results of the first molecular characterisation of grapevine viruses in Bosnia and Herzegovina.

Genetic Structure and Molecular Variability of Grapevine Fanleaf Virus in Sicily

“…This result may be favored by the establishment of GFLV in the different vine-growing Sicilian areas due to the continuous exchange of GFLV-infected propagation plant material and the possible presence of its vector (X. index), as already reported in Italy [29]. In addition, it is possible to hypothesize that cultivars that showed a higher incidence of infection are more susceptible to GFLV infection than others, as was similarly found in autochthonous cultivars in another GFLV study [34].…”

“…Reverse-transcription polymerase chain reaction (RT-PCR) assays were carried out on samples that resulted in being positive for GFLV/ArMV by DAS-ELISA and "Grillo" and "Moscato" samples (Table 1). In detail, the GFLV coat protein (CP) and ArMV polyprotein genes were amplified by end point RT-PCR, using the EV00N1/EV00N3 [21] and ArMV 2B/ArMV 2BR [34] primer pairs, respectively (Table 2). The specific primers used for GFLV amplification designed by Vigne and co-workers [21] were modified in order to delete the sequence of the restriction enzyme XbaI.…”

“…RT-PCR was carried out in a MultiGene OptiMax thermal cycler (Labnet International Inc., Edison, NJ, USA). The cycling conditions were as follows: reverse transcription at 42 • C for 50 min, initial denaturation at 95 • C for 5 min, followed by 40 cycles of 95 • C for 30 s, 54 • C (ArMV) [34]-55 • C (GFLV) [21] for 1 min and 72 • C for 90 s, and a final elongation step at 72 • C for 10 min. Total RNAs derived from grapevine infected by GFLV and ArMV (Agritest srl, Valenzano, Italy) were used as positive control, while molecular-grade water and total RNA extracted from healthy grapevine plant (Agritest srl, Valenzano, Italy) were used as negative controls.…”

Viruses Infecting Trees and Herbs That Produce Edible Fleshy Fruits with a Prominent Value in the Global Market: An Evolutionary Perspective

“…In this regard, grapevine leafroll disease is an excellent example of a multiple viral infection, as it is caused by the association of up to 11 grapevine leafrollassociated viruses [108]. In fact, an extensive survey of five major grapevine viruses indicated a putative recombination event for the grapevine leafroll-associated virus 3 (GLRaV-3; Ampelovirus, Closteroviridae) [109]. On the other hand, Hanssen et al found that PepMV recombinants frequently occur in mixed infections under natural conditions [110], showing that coexistence of viruses is a prerequisite for recombination events.…”

“…China, France, United States, Spain, and Chile are the main countries with grapevine producing areas [45]. V. vinifera is a host for at least 90 viruses (Figure 4A; Supplementary Table S4), but only a few of them are of economic importance in the world, such as grapevine fanleaf virus (GFLV; Nepovirus, Secoviridae) and several species within the Closteroviridae family designated as grapevine leafroll-associated viruses (Supplementary Table S4) [108,109,121,122]. GFLV is transmitted by Xiphinera index, an ectoparasite that feeds on plant roots and can resist long periods in the soil.…”