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1.
Sixteen Plum pox virus (PPV) isolates from several stone fruit cultivars, host species, orchards and geographical areas of Bosnia 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 Rsa I and Dde I. Four PPV isolates were identified as PPV-M by serology and PCR; eight isolates were identified as PPV-D based on PCR–RFLP on both genomic regions, but were not recognized by the D-specific MAb4DG5. Four isolates from plum were identified as natural D/M recombinants (PPV-Rec), based on conflicting results of CP and P3-6K1 typing. To investigate the genetic diversity of Bosnian PPV isolates in more detail, five isolates (three PPV-Rec, one PPV-M and one PPV-D) were partially sequenced in the region spanning the 3' terminal part of the NIb gene and the 5'-terminal part of the CP gene, corresponding to nucleotides 8056–8884. Nucleotide sequence alignment of recombinant isolates showed that they were closely related at the molecular level to previously characterized recombinants from other European countries, and shared the same recombination break point in the 3' terminal part of the NIb gene. This is the first report of naturally infected Prunus trees with PPV-M, PPV-D and PPV-Rec in Bosnia and Herzegovina. The high variability of the Bosnian PPV isolates fits with the presence of this virus in the country over a long period. 相似文献
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When the first foci of sharka were discovered in Puglia region (south-east Italy) in the late 1980s, the regional agricultural authorities launched a programme for Plum pox virus (PPV) monitoring and disease eradication. The infecting virus strain was identified as PPV-D. From 1989 to 1993, a strong eradication campaign was successfully carried out involving 13 plum and 2 apricot orchards with different levels of infection. During 1994–2000, besides plum, apricot and peach, monitoring was extended to sweet cherry. At that time, surveys and testing did not reveal any new PPV focus, but the eradication of infected trees continued in a couple of orchards. In 2001–05, particular attention was paid to peach, as devastating PPV-M outbreaks had developed in other areas of the country. A new PPV focus was found in apricot, caused by PPV-Rec, which was promptly eradicated. In the following two years, surveys in the once infected orchard and surrounding peach plantings did not detect any virus spread. The endeavour has taken 15 years making this PPV monitoring and eradication programme the longest in Italy. Its overall results indicate that the fruit tree industry in Puglia region can now be regarded as essentially PPV-free. 相似文献
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An Italian isolate of plum pox potyvirus (PPV) from apricot, Ispave 17, was used as antigen for production of monoclonal antibodies. Six clones secreting specific antibodies to PPV were obtained. All these monoclonal antibodies were used to test a collection of different Italian PPV isolates, collected from plum, apricot and peach orchards, and other European isolates (including PPV-D and PPV-M serotypes), using DAS-ELISA, SDS-PAGE, western blot and GIEM. In western blot analysis, the PPV-M and PPV-D coat protein, detected directly from crude peach GF305 extracts, showed different electrophoretic mobility, the coat protein of PPV-M being slightly larger than that of PPV-D. ELISA tests, performed with fixed dilutions of antibodies and limiting dilutions of clarified samples, showed with some monoclonal antibodies a marked difference between PPV-M and PPV-D strains, at ratios greater than 1:40 (w/v). Also in GIEM some monoclonal antibodies gave a good labelling reaction only with PPV-D serotype. With the help of this differentiation, it was found that all Italian isolates tested were of the D serotype and none of the severe M strain of PPV, which has not been reported in Italy. 相似文献
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Kensaku Maejima Misako Himeno Osamu Netsu Kazuya Ishikawa Tetsuya Yoshida Naoko Fujita Masayoshi Hashimoto Ken Komatsu Yasuyuki Yamaji Shigetou Namba 《Journal of General Plant Pathology》2014,80(2):176-183
Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains. 相似文献
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Epidemiology of sharka disease in France 总被引：1，自引：0，他引：1
Plum pox virus was first detected in France in the 1960s. Both PPV-D and PPV-M strains are present but epidemics related to the PPV-M strain detected in the late 1980s are the most problematic. The two PPV strains have unequal distributions in peach and apricot orchards and different prevalences. More than 20 different aphid species have been identified as vectors of PPV but most of them do not colonize Prunus species. Thus, aphids involved in the spread of PPV in orchards are essentially visiting aphids. The main sources of inoculum for the vectors are leaves and fruits of infected stone-fruit trees. Spontaneous, wild and ornamental Prunus species such as Prunus dulcis , P. spinosa or P. pissardii are susceptible to PPV isolates found in France but their role as a reservoir in sharka epidemics is probably negligible. The disease spreads rapidly in orchards but the rate of progression may vary according to the identity of the PPV strain and the Prunus species. Analysis of spatial patterns of disease has shown that secondary spread by aphids frequently occurs over short distances in the orchards (aggregated patterns) but also that dissemination at longer distances (of several hundred metres) is a common event. 相似文献
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Arben Myrta Donato Boscia Oriana Potere Maria Kölber Maria Németh Biagio Di Terlizzi Mariano Cambra Vito Savino 《European journal of plant pathology / European Foundation for Plant Pathology》2001,107(8):845-848
A large-scale serological characterisation of Plum pox virus (PPV) isolates was carried out with 19 monoclonal antibodies (MAbs), including the universal MAb5B and the following strain-specific MAbs: AL (specific to PPV-M), 4DG5 (specific to PPV-D), TUV and AC (specific to PPV-C), and EA24 (specific to PPV-EA). The study involved 108 PPV isolates of different geographical origin (Albania, Bulgaria, Cyprus, Czech Republic, Egypt, France, Germany, Greece, Italy, Hungary, Moldova, Romania, Slovakia, Spain, Turkey and Yugoslavia) and hosts (almond, apricot, peach, plum and cherry). The inter- and intra-strain serological relationships of PPV isolates were evaluated by DASI-ELISA. High serological variability was detected, not only between strains, but also among isolates of the same strain. Computer-assisted analysis of serological data support the hypothesis of the existence of two distinct subclusters, denoted PPV-M1 and PPV-M2, which seem to prevail in Mediterranean and Eastern–Central European countries, respectively. 相似文献
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Plum pox virus (PPV) strain D is globally distributed and causes serious losses in stone fruits in over 40 countries. Here, full-length genomic sequences were analysed for 44 PPV-D isolates from all regions of Turkey, together with partial sequences for a larger number of isolates. PPV-D isolates from Turkey are similar to other PPV-D isolates in all major genomic features. However, the majority of Turkish PPV-D isolates form separate phylogenetic clusters from all other isolates and show a geographical clustering tendency, suggestive of limited movement between regions. In particular, PPV-D isolates from Thrace and Central Anatolia formed a monophyletic sister cluster to the cluster that includes all previously known PPV-D isolates. Two isolates with strong evidence of recombination with the PPV-T strain were identified, together with two isolates with weaker evidence for intra-D strain recombination. The genetic diversity of PPV-D was found to be particularly high in Turkey (0.017 ± 0.001%), close to that observed for PPV-D world diversity once the over-represented isolates from Japan, the USA and Canada have been excluded (0.020 ± 0.001%). Taken together, these results suggest a long and largely isolated evolutionary history of PPV-D in Turkey and further extend knowledge of the diversity of this highly successful strain. The high diversity of PPV-D in Turkey, together with the basal phylogenetic position of Turkish isolates, are compatible with a hypothesis making Turkey the centre of origin of the D strain. 相似文献
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Plant RNA viruses have a high genetic variation potential due to the absence of proofreading activity in their RNA replicase. In addition to mutation, recombination is generally thought to be an important source of variability. Both evolutionary processes have contributed to the diversity of Plum pox virus (PPV). There are now six recognized subgroups, strains or serotypes of PPV (D, M, Rec, EA, C and W). Isolates belonging to the PPV-Rec subgroup are derived from RNA recombination between PPV-D and PPV-M and occur frequently in various central and eastern European countries. The divergent isolate W3174 is a new and distinct strain of PPV, identified as PPV-W. It is quite conceivable that, with time, other groups will be defined and that the present classification will need revision to accommodate additional PPV variability. 相似文献
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Kensaku Maejima Hideo Hoshi Masayoshi Hashimoto Misako Himeno Takeshi Kawanishi Ken Komatsu Yasuyuki Yamaji Hiroshi Hamamoto Shigetou Namba 《Journal of General Plant Pathology》2010,76(3):229-231
For the first time, plum pox virus (PPV) has been detected in commercial Japanese apricot (Prunus mume) trees in Tokyo, Japan. These trees had ringspot or mottle on leaves, color breaking of petals and, occasionally, mild ringspots
and malformation on fruits. The virus was identified based on the morphology of virus particles, serology, and RT-PCR. The
amplified nucleotide fragment shared 100% identity with a partial coat protein gene of PPV-D isolates. 相似文献
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Schneider WL Damsteegt VD Gildow FE Stone AL Sherman DJ Levy LE Mavrodieva V Richwine N Welliver R Luster DG 《Phytopathology》2011,101(5):627-636
Plum pox virus (PPV) was identified in Pennsylvania in 1999. The outbreak was limited to a four-county region in southern Pennsylvania. Initial serological and molecular characterization indicated that the isolates in Pennsylvania belong to the D strain of PPV. The Pennsylvania isolates were characterized by sequence analysis, electron microscopy, host range, and vector transmission to determine how these isolates related to their previously studied European counterparts. Genetically, Pennsylvania (PPV-Penn) isolates were more closely related to each other than to any other PPV-D strains, and isolates from the United States, Canada, and Chile were more closely related to each other than to European isolates. The PPV-Penn isolates exist as two clades, suggesting the possibility of multiple introductions. Electron microscopy analysis of PPV-Penn isolates, including cytopathological studies, indicated that the virions were similar to other Potyvirus spp. PPV-Penn isolates had a herbaceous host range similar to that of European D isolates. There were distinct differences in the transmission efficiencies of the two PPV-Penn isolates using Myzus persicae and Aphis spiraecola as vectors; however, both PPV-Penn isolates were transmitted by M. persicae more efficiently than a European D isolate but less efficiently than a European M isolate. 相似文献
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Epidemiology of sharka disease in Spain 总被引：1，自引：0，他引：1
M. Cambra N. Capote M. A. Cambra G. Llácer P. Botella A. López-Quílez 《EPPO Bulletin》2006,36(2):271-275
PPV was first detected in Spain in 1984 in Japanese plum ( Prunus salicina Lindl) cv. Red Beaut and spread very quickly to other Japanese and European plums and apricot cultivars but left peach cultivars unaffected. In the years following the detection of PPV, the predominant aphid species visiting Prunus orchards in Mediterranean areas were Aphis gossypii followed by Aphis spiraecola , the latter being the main aphid species found at present. Both species are considered to be the main vectors of PPV in Spanish early Prunus growing areas. Spatial analysis of the spread of PPV-D in Japanese plum and apricot trees confirmed the lack of significant association between immediately adjacent trees. The observed spatial pattern of sharka suggests a lack of movement of PPV-viruliferous aphid vectors to immediately adjacent trees and indicates their preferential movement to trees several tree spaces away. PPV-D is the only type currently present in Spain, with the exception of a PPV-M outbreak that was detected in and successfully eradicated from Aragón in 2002. The short-distance spread of PPV-M infection occurred as far as 12 m along the rows of peach trees. However, PPV-D has not been observed to spread through peach cultivars, despite being grown in the vicinity of heavily infected plots of apricot or Japanese plum trees. 相似文献
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Gürcan Kahraman Teber Saffet Akbulut Mikail Çağlayan Kadriye 《European journal of plant pathology / European Foundation for Plant Pathology》2021,161(2):453-461
European Journal of Plant Pathology - Plum pox virus strain Recombinant (PPV-Rec) is hypothetically considered as homologous recombinant between strains PPV-M and PPV-D. The nucleotide position... 相似文献
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Miroslav Glasa Véronique Marie-Jeanne Gérard Labonne Zdeno Šubr Otakar Kúdela Jean-Bernard Quiot 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(9):843-853
The isolate BOR-3, collected in Slovakia in 1996, was recently identified as a natural recombinant between an M and D type of Plum pox virus (PPV). Biological assays demonstrated its capacity to be aphid- and graft-transmitted to various Prunus spp. hosts. A study was carried out to determine the further presence of PPV recombinants in two epidemiologically distinct areas – Slovakia and France. Tools based on PPV-M and D subgroup typing, targeting P3–6K1, CI and CP regions of the PPV genome were used for recombinant identification. Closely related recombinant variants were detected in different Prunus spp. during a survey conducted in Slovakia in 2001, but not within a set of selected PPV isolates from France collected between 1985 and 2001. Sequence analysis of the (Cter)NIb–(Nter)CP region of 10 recombinant isolates from Slovakia showed their high homology, reaching more than 98%. All the recombinant isolates shared the same recombination breakpoint situated in the C terminus of the NIb gene. Our study demonstrates that the PPV recombinants are viable and competitive with conventional PPV-M and D isolates. The present work indicates that the occurrence of recombinants within PPV isolates might be more common than previously assumed. 相似文献
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M. Hassan G. Gomez V. Pallás A. Myrta P. Rysanek 《European journal of plant pathology / European Foundation for Plant Pathology》2009,124(2):363-368
In this paper, we report a large-scale survey for the incidence of Peach latent mosaic viroid (PLMVd) and Hop stunt viroid (HSVd) in stone fruit collections and commercial orchards in the Czech Republic. From the 645 samples analysed, PLMVd was
detected in 80 (26.6%) of peaches and the HSVd in 3 (1.3%) of apricot and 1 (0.33%) of peach trees. Sixty-seven accession
of peach (44.6%) from the Czech Clonal GeneBank were infected by PLMVd. In addition, we used naturally infected trees to standardise
the simultaneous detection of PLMVd and HSVd plus host mRNA as the control by means of one-step multiplex RTC-PCR. Eleven
PLMVd and two HSVd isolates were sequenced and analysed. All the PLMVd variants were highly homologous (97–100%) to previously
reported PLMVd variants from Tunisian peach and almond trees, and clustered together in the previously reported phylogenetic
group III. The HSVd variants obtained from apricot and peach trees were included in the previously proposed recombinant group
PH/cit3. 相似文献
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Jürgen Köhl Belia Groenenboom-de Haas Helen Goossen-van de Geijn Adrianus Speksnijder Pieter Kastelein Sybren de Hoog Bert Gerrits van den Ende 《European journal of plant pathology / European Foundation for Plant Pathology》2009,124(1):151-162
Stemphylium vesicarium (teleomorph: Pleospora herbarum) is the causal agent of brown spot disease in pear. The species is also able to cause disease in asparagus, onion and other
crops. Saprophytic growth of the fungus on plant debris is common. The objective of this study was to investigate whether
isolates of S. vesicarium from different hosts can be pathogenic to pear. More than hundred isolates of Stemphylium spp. were obtained from infected pear fruits, dead pear leaves, dead grass leaves present in pear orchard lawns as well as
from necrotic leaf parts of asparagus and onion. Only isolates originating from pear orchards, including isolates from dead
grass leaves, were pathogenic on pear leaves or fruits in bioassays. Non-pathogenic isolates were also present in pear orchards.
Stemphylium vesicarium from asparagus or onion, with one exception, were not pathogenic to pear. Analysis of the genetic variation between isolates
using Amplified Fragment Length Polymorphism (AFLP) showed significant concordance with host plants. Isolates from asparagus
or onion belonged to clusters separate from the cluster with isolates from pear or grass leaves collected in pear orchards.
Multilocus sequencing of a subset of isolates showed that such isolates were similar to S. vesicarium. 相似文献