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The Korean Society of Plant Pathology (한국식물병리학회)
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Update on Distribution and Genetic Variability of Plum pox virus Strains in Bulgaria

  • Received : 2018.09.17
  • Accepted : 2018.12.02
  • Published : 2019.06.01
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Abstract

Field surveys for Plum pox virus (PPV) infection were conducted in stone fruit orchards all over Bulgaria. In total, 1168 out of 3020 leaf samples from cultivated Prunus spp. and wildly growing P. cerasifera trees reacted positive for PPV in DASI-ELISA with the universal monoclonal antibody (MAb) 5B. Further ELISA analyses showed that 987 and 127 isolates belonged to PPV-M and PPV-D serotypes, respectively. The plum and P. cerasifera showed 82.0% and 50.5% levels of infection, respectively followed by the peach (40.0%) and the apricot (32.0%). Five hundred fifty one PPV isolates were further typed by IC-RT-PCR with PPV-Rec, -M and -D-specific primers, targeting (Cter)NIb-(Nter) CP genome region, as 125 isolates were sequenced. The results revealed the presence of PPV-Rec, PPV-M and PPV-D and mixed infections of these strains. PPV-Rec was the most prevalent strain (49.0%), followed by PPV-M (40.1%), while PPV-D was the less spread strain (8.2%). PPV-Rec was the most common strain in plums, including the eight "old-aged" trees from the region of the first Sharka discovery. PPV-M was the most prevalent strain in peach and apricot. Phylogenetic analyses on (Cter)NIb-(Nter)CP of the isolates were performed. PPV-Rec isolates formed a homogeneous group, while PPV-M isolates split into PPV-Ma and PPV-Mb subgroups. Five separated clades were formed by the analyzed PPV-D isolates. Nucleotide sequences of the partial CP coding region of the analyzed isolates revealed a slightly higher intra-strain genetic variability in PPV-Rec and PPV-M isolates, while that of PPV-D strain isolates was higher from the reported for these strains.

Keywords

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Fig. 1. Phylogenetic tree of Plum pox virus strain Rec isolates from Bulgaria reconstructed from (Cter)NIb-(Nter)CP genomic region. The scale represents a distance of 0.05 substitutions per site. The tree was reconstructed by neighbour joining using the Kamura-2 parameter model of evolution. Only bootstrap values > 70% are shown. The isolates sequenced in this study are boldface. After the name of the isolate the original host is shown as plplum, pe-peach, ap-apricot and p.c-P. cerasifera.

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Fig. 2. Phylogenetic tree of Plum pox virus strain M isolates from Bulgaria reconstructed from (Cter)NIb-(Nter) CP genomic region. The scale represents a distance of 0.05 substitutions per site. The tree was reconstructed by neighbour joining using the Kamura-2 parameter model of evolution. Only bootstrap values > 70% are shown. The isolates sequenced in this study are boldface. After the name of the isolate the original host is shown as pl-plum, pe-peach, ap-apricot and p.c-P. cerasifera.

E1PPBG_2019_v35n3_243_f0003.png 이미지

Fig. 3. Phylogenetic tree of Plum pox virus strain D isolates from Bulgaria reconstructed from (Cter)NIb-(Nter)CP genomic region. The scale represents a distance of 0.05 substitutions per site. The tree was reconstructed by neighbour joining using the Kamura-2 parameter model of evolution. Only bootstrap values > 70% are shown. The isolates sequenced in this study are boldface. After the name of the isolate the original host is shown as pl-plum, pe-peach, apapricot and p.c-P. cerasifera.

Table 1. List of PPV isolates characterized in this study or available in GenBank

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Table 1. Continued

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Table 2. Incidence of PPV strains in particular Prunus spp. in Bulgaria

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Table 3. Mean nucleotide divergence levels in the partial coat protein gene of PPV-Rec isolates

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