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Genetic Diversity of Seven Strawberry mottle virus Isolates in Poland

  • Received : 2018.12.31
  • Accepted : 2019.04.03
  • Published : 2019.08.01

Abstract

The studies on detection of the Strawberry mottle virus (SMoV) have been conducted in Poland for breeding programme purpose and for producers of strawberry plant material. Leaf samples collected from infected strawberry plants were grafted on Fragaria sp. Indicators which were maintained in greenhouse for further study. Seven Fragaria vesca var. semperflorens 'Alpine' indicators infected by SMoV were used for the study aimed on molecular characterization of virus isolates. Partial RNA2 was amplified from total nucleic acids using the RT-PCR method. The obtained amplicons separately digested with BfaI, FauI, HaeIII, HincI, and TaqI enzymes showed different restriction profiles. The nucleotide sequences analysis of RNA2 fragment confirmed the genetic diversity of the SMoV isolates as their similarity ranged from 94.7 to 100%. Polish isolates shared 75.7-99.2% identity with sequence of the virus strains from the Czech Republic, the Netherlands, and Canada. Phylogenetic analysis resulted in grouping of the isolates found in Poland together with one of the Czech strain whereas two other from the Czech and the strains from the Netherlands and Canada created the separate cluster.

Keywords

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Fig. 1. Polyacrylamide gel showing the RFLP patterns of RNA2 fragment of the Strawberry mottle virus isolates amplified with 2SMoV1108F-2SMoV2386R primers digested with BfaI, FauI, HaeIII, HincI, and TaqI restriction enzymes of strawberry samples collected in 2015 in Bulgaria and Poland. Lanes: L – Thermo Scientific GeneRuler 100 bp Plus DNA Ladder, fragment sizes in base pairs (from top to bottom): 3000, 2000, 1500, 1200, 1000, 900, 800, 700, 600, 500, 400, 300, 200, 100. Samples: 1. Unkn-1108, 2. Granda-1108, 3. Markat-1108, 4. Pink-1108, 5. karkas-1108, 6. Pegat-1108, 7. Granat-1108.

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Fig. 2. Phylogenic analysis of the RNA2 fragment sequences of the SMoV isolates detected in Poland and nine representative SMoV strains. The SMoV isolates characterized during this study are indicated in bold. Phylogenetic tree was constructed by the maximum likelihood algorithm (MEGA 5.2) and the Kimura 3-parameter nucleotide substitution model in using 1,000 bootstrap replicates. Only bootstrap values higher than 60 are shown. The bar indicates the genetic distance as Kimura units. RNA2 sequence of Satsuma dwarf virus (GenBank accession number: AB009959) was used as the external group.

Table 1. Diversity of the RFLP profiles after digestion of RNA2 fragment of SMoV isolates by BfaI, FauI, HincI, TaqI, and HaeIII enzymes. Designation by A or B or C letters indicates the presence of different restriction patterns for the specific enzyme

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