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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
Full text linkTissue-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
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 report of Little cherry virus 1 in cherry, plum, almond and peach in Italy
No full textLittle cherry disease (LChD) is a widespread disorder of
ornamental, sweet and sour cherries. In sensitive cultivars,
it results in the production of small, pale-coloured fruits
with reduced sugar content and in the premature reddening
or bronzing of the leaves. Little cherry virus-1 (LChV-
1) and Little cherry virus 2 (LChV-2), both members of the
family Closteroviridae, are associated with this disease, but
often induce symptomless infection. Assays for both viruses
were made during a survey in 2006 and 2007 of the sanitary
status of fruit trees in Apulia (southern Italy). Samples
were collected in different commercial orchards from 22
sweet cherry, 13 plum, five almond, five peach, and two
apricot trees. Total nucleic acids (TNA) were extracted
from the leaves as described by Foissac et al. (2001) and
used as template for Superscript III one-step RT-PCR with
Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA,
USA) using primer sets specific for LChV-1 or LChV-2
(Rott and Jelkmann, 2001). Whereas all samples were negative
for LChV-2, a 419 bp fragment corresponding to part
of the 3' non-translated region of LChV-1 RNA was amplified
from five cherry, four plum, one almond and one
peach tree samples. These results were obtained in several
independent experiments. Trees of both native and imported
cultivars were infected but LChV-1 was not associated
with any particular field symptoms. To our knowledge,
this is the first report of LChV-1 in Italy and of its
natural occurrence in plum, almond and peach
Detection of three closteroviruses in stone fruit trees by multiplex assays
No full textSimultaneous detection of Plum bark necrosis stem pitting-
associated virus (PBNSPaV), Little cherry virus 1
(LChV-1) and Little cherry virus 2 (LChV-2) was
achieved with a quadruplex one-step RT-PCR, and a hybridization
test with a multi-riboprobe, apparently representing
the first successful attempt of concomitant identification
of viruses of the family Closteroviridae in stone
fruit trees. Quadruplex RT-PCR detected double (natural
and artificial) and triple (artificial) infections in all samples
tested, amplifying also a plant mRNA as an internal
control. Standard and quadruplex RT-PCR were comparable
in terms of detection limits and specificity. Molecular
hybridization with a multi-riboprobe (‘clotri’) containing
partial sequences of PBNSPaV, LChV-1 and LChV-2,
allowed the detection of each of the three viruses in
Prunus hosts. The sensitivity of single riboprobes and
“clotri” was comparable
Monitoring of Plum bark necrosis stem pitting-associated virus and molecular characterization of some isolates
No full textDifferent Prunus species from Italy, Serbia, Turkey
and Egypt were assayed by one-step RT-PCR and nested
PCR for the presence of Plum bark necrosis stem pittingassociated
virus (PBNSPaV). This virus had a fairly high
incidence and was monitored throughout the year by
IC-RT-PCR and ELISA. It was detected during all seasons
by both techniques but most easily in spring in the
new flushes of vegetation. Six PBNSPaV isolates were
characterized by partial sequencing of three genomic regions,
e.g. HSP70, coat protein and ORF4. High nucleotide
similarity was found between all isolates from
different geographic and host origins, suggesting either
a relatively recent origin or the presence of unidentified
constraint on variation
Viruses and viroids of stone fruits in Egypt. Short communication
No full textField surveys were carried out to assess the sanitary
status of stone fruits in Egypt. A total of 716 samples
was tested by ELISA for Prunus necrotic ring spot virus
(PNRSV), Plum pox virus (PPV), Prune dwarf virus
(PDV), Apple chlorotic leaf spot virus (ACLSV) and Apple
mosaic virus (ApMV). The viruses most frequently
detected were PNRSV (57%) and PPV (41%); PDV
and ACLSV were found in 3% of samples and ApMV
was not detected. Plum bark necrosis stem pitting-associated
virus (PBNSPaV) and Apricot latent virus (ApLV)
were detected for the first time in Egypt by using RTPCR.
Of 693 samples tested by tissue-imprint hybridization
(TIH), 101 (15%) were found to be infected by either
Peach latent mosaic viroid (PLMVd) or Hop stunt
viroid (HSVd
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