67 research outputs found
Diversity of Plum Pox Virus isolates in Bosnia and Herzegovina
No full textSixteen Plum pox virus (PPV) isolates from several stone fruit cultivars, host species, orchards and geographical areas ofBosnia and Herzegovina were selected for typing, using serotype-specific monoclonal antibodies (MAbs) and PCR–RFLP,targeting the 3′ terminal region of the coat protein (CP) and P3-6K1 with restriction enzymes RsaI and DdeI. Four PPVisolates were identified as PPV-M by serology and PCR; eight isolates were identified as PPV-D based on PCR–RFLP onboth genomic regions, but were not recognized by the D-specific MAb4DG5. Four isolates from plum were identified asnatural D/M recombinants (PPV-Rec), based on conflicting results of CP and P3-6K1 typing. To investigate the geneticdiversity of Bosnian PPV isolates in more detail, five isolates (three PPV-Rec, one PPV-M and one PPV-D) were partiallysequenced in the region spanning the 3′ terminal part of the NIb gene and the 5′-terminal part of the CP gene, corre-sponding to nucleotides 8056–8884. Nucleotide sequence alignment of recombinant isolates showed that they wereclosely related at the molecular level to previously characterized recombinants from other European countries, andshared the same recombination break point in the 3′ terminal part of the NIb gene. This is the first report of naturallyinfected Prunus trees with PPV-M, PPV-D and PPV-Rec in Bosnia and Herzegovina. The high variability of the BosnianPPV isolates fits with the presence of this virus in the country over a long perio
First record of Peach latent mosaic viroid and Hop stunt viroid in Bosnia and Herzegovina
No full textDuring a survey for the evaluation of the sanitary status
of the stone fruit industry of Bosnia and Herzegovina,
410 samples were collected from 33 commercial orchards
and 2 nurseries, and tested for the presence of
Peach latent mosaic viroid (PLMVd) and Hop stunt viroid
(HSVd). Samples were from 230 plants of plum, 77
of peach, 65 of cherry, 11 of apricot, and 27 of other
Prunus species (myrobalan and blackthorn) growing in
the surroundings of Banja Luka, Gradacac, Sarajevo, or
Mostar. All samples were tested by tissue-print hybridization
(Pallás et al., 2003), by pressing the freshly
cut end of a leaf petiole onto a Hybond N+ nylon membrane
and hybridizing it at 55°C with SP6 and T7 RNA
polymerase-generated full-length digoxygenin-labelled
viroidal cRNA probes (Shamloul et al., 1995; Astruc et
al., 1996). About 10% of the samples (43 of 410) were
found to be infected by viroids. In particular, PLMVd
was detected in 39 plants of peach, and HSVd in two of
apricot and two of plum. Positive samples were from
both native and imported cultivars. Interestingly, among
PLMVd-infected peaches, two were seedlings. Whether
the infection had come via pruning tools or via seeds remains
to be established. This report is the first record of
PLMVd and HSVd in Bosnia and Herzegovina
Occurrence of stone fruit viroids in Bosnia and Herzegovina
No full textTissue-imprint hybridization (TIH) assays were used to determine the occurrence and incidence of Peach
latent mosaic viroid (PLMVd) and Hop stunt viroid (HSVd) in stone fruit trees in Bosnia and Herzegovina. Our
collections included trees of plum, peach, cherries, apricot, myrobalan and blackthorn from 33 commercial orchards
and 2 nurseries, in the areas of Banja Luka, Gradacac, Sarajevo and Mostar. Of the 410 trees assayed, 44 (11%)
tested positive by TIH assays. PLMVd was detected in 39 peach trees, including two old (seed grown) vineyard peach
trees (Prunus persica subsp. vulgaris). Tests for HSVd were positive in 3 apricot and 2 plum trees. PLMVd was
widely distributed throughout the country. In contrast, HSVd was found only in the northern part of the country.
Both native and imported cultivars of Prunus were infected. This is the first record of PLMVd and HSVd in Bosnia
and Herzegovina. In a separate experiment, peach trees with PLMVd were monitored in the autumn, winter and
spring seasons, with tissue imprints of leaf petioles, dormant cuttings and forced sprouts from dormant cuttings.
Irrespective of the tissues assayed, nearly all samples tested positive for PLMVd
Viruses of stone fruits in Bosnia and Herzegovina
No full textDuring spring 2004, field surveys were carried out in the main stone fruit growing areas of Bosnia and Herzegovina to assess the sanitary status of commercial orchards, nurseries, home gardens and bordering trees. A total of 1,080 samples: 592 plums, 244 peaches, 149 cherries, 68 apricots and 27 other Prunus species (myrobalan and blackthorn) were tested by ELISA for the presence of Plum pox virus (PPV), Prunus necrotic ring spot virus (PNRSV), Prune dwarf virus (PDV), Apple chlorotic leaf spot virus (ACLSV) and Apple mosaic virus (ApMV). The overall average infection level was 36%. The most infected species was peach (63%) and the least apricot (16%). The most frequent virus was PPV (47%), followed by PNRSV (37%), PDV (26%) and ACLSV (4%). No ApMV was detected. The infection rate in commercial orchards was 41% and in gardens and bordering trees 38%. Virus infection in nurseries (21%) indicated their potential role in spread of virus diseases into new plantings. Stone fruit viruses were detected in local and imported cultivars and found widely distributed throughout the country. Our study revealed a deteriorated sanitary status for the local stone fruit industry
High-Throughput Sequencing: Advantages Beyond Virus Identification
No full textThe application of high-throughput sequencing (HTS) in plant virology research had an immense influence on our perspective of disease etiology. The technology opened a new avenue of exploration that is unbiased and at an unparalleled level of sensitivity. Initially, HTS was used for virus discovery with the greatest success up until now being the discovery of Grapevine red blotch-associated virus (GRBaV) and Grapevine Pinot gris virus (GPGV). Most of the early studies constructed HTS libraries from either an enriched dsRNA extract or the small RNA fraction, but other nucleic acids have also been used. The expansion of virus databases and improved bioinformatic tools would suggest that HTS will be implemented as a sensitive virus detection tool more routinely. The discovery of the complex grapevine virome challenges the concept of what can be considered as a healthy vine although offering new opportunities to the implementation of certification schemes and exchanges of plant propagation materials.
In this chapter the history of HTS in grapevine virus research is reviewed followed by the exploration of the different strategies used for the discovery of new viruses and new variants of known viruses. The concept of a “background” virome is introduced and explored within the context of its interaction with the host and the impact this might have on certification of healthy plant material. The chapter is concluded with some thoughts on the future of HTS in grapevine virus research
First report of grapevine associated jivivirus 1 infecting grapevines in Brazil.
No full textGrapevines can host up to 86 virus species, some of which affect plant vigor, production and fruit quality (Fuchs, 2020). In 2014, a Vitis vinifera cv. Semillon vine showing yellow speckles and mild leafroll symptoms in Bento Gonçalves, RS, Brazil, was investigated for viruses (Silva et al., 2017), resulting in the detection of grapevine enamovirus 1, grapevine yellow speckle viroid 1 and hop stunt viroid
Viruses of sweet and sour cherry in Serbia
No full textOne hundred twenty-five trees (each of a different
cultivar) of sour and sweet cherry from two large varietal
collections in Serbia were visually inspected for
virus symptoms and tested for the presence of cherry
viruses by ELISA, herbaceous host assays, graft-indexing
on P. serrulata cv. Kwanzan, and RT-PCR. All samples
were tested by ELISA for Prunus necrotic ring spot
virus (PNRSV), Prune dwarf virus (PDV), Apple chlorotic
leaf spot virus (ACLSV), Apple mosaic virus (ApMV),
and Plum pox virus (PPV). The overall detection of
PDV, PNRSV, and ACLSV was 63%. Additional
ELISA tests were done on 80 trees for Arabis mosaic
virus (ArMV), Cherry leaf roll virus (CLRV), Strawberry
latent ring spot virus (SLRSV), Petunia asteroid mosaic
virus (PetAMV), Raspberry ringspot virus (RpRSV),
Tomato black ring virus (TBRV), Tobacco mosaic virus
(TMV), and Tomato ringspot virus (ToRSV). In these
tests, one tree tested positive for PetAMV. RT-PCR testing
of 44 trees detected another five viruses: Cherry
green ring mottle virus (CGRMV), Cherry necrotic rusty
mottle virus (CNRMV), Cherry virus A (CVA), European
rusty mottle associated virus (ERMaV) and Plum
bark necrosis and stem pitting-associated virus (PBNSPaV),
but not Cherry mottle leaf virus (CMLV). In
graft-indexing tests on Kwanzan with all 125 trees, samples
from 38 trees induced symptoms of necrotic crook
disease (causal agent unknown). Viruses reported for
the first time in Serbia were CGRMV, CNRMV, CVA,
ERMaV, PBNSPaV, and PetAMV
MOLECULAR VARIABILITY OF APPLE CHLOROTIC LEAF SPOT VIRUS IN DIFFERENT HOSTS AND GEOGRAPHICAL REGIONS
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