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Genetic diversity of Czech ‘Candidatus Phytoplasma mali’ strains based on multilocus gene analyses

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Abstract

In several European countries apple trees are affected by apple proliferation disease, which is usually associated with the presence of ‘Candidatus Phytoplasma mali’. During 2010, samples from several apple trees displaying proliferation symptoms were collected throughout the Czech Republic to verify identity of phytoplasmas detected in association with the disease. The majority of the 74 apple trees examined using molecular tools were positive for ‘Ca. P. mali’ presence. The 16S–23S ribosomal genes, the ribosomal protein genes and the nitroreductase and rhodonase like genes were then studied to verify phytoplasma strain variability on multigenic bases. Two RFLP profiles and correspondingly two genetic lineages were found in the PCR-amplified fragments covering the 16S–23S rDNA spacer region. ‘Ca. P. mali’ strains belonging to rpX-A subgroup were identified in the majority of the apple tree sampled, whereas phytoplasmas belonging to the rpX-B subgroup were distributed sporadically. The apple proliferation subtypes AP-15 and AT-2 exhibited nearly equal occurrence; the AT-1 subtype and a mixture of the two or all three of the AP subtypes were infrequently found. The PCR/RFLP results were confirmed by nucleotide sequence analyses of selected ‘Ca. P. mali’ strains.

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Acknowledgments

The work was supported by grant of Ministry of Education, Youth and Sports of the Czech Republic No. OC09021 (Cost Action 864), and No. LD12074 (Cost Action FA0807) and the institutional support RVO:60077344. The authors are deeply indebted to P.A. Bianco (Università degli Studi di Milano, Italy) for valuable advices, to A. Matyášová (BC ASCR v.v.i. IPMB, České Budějovice, Czech Republic) for excellent technical assistance and to all workers of State Phytosanitary Service of the Czech Republic for skilled samples collection.

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Authors and Affiliations

  1. Department of Plant Virology, Biology Centre of the ASCR, v.v.i., Institute of Plant Molecular Biology, Branišovská 31, 370 05, České Budějovice, Czech Republic

    Jana Fránová

  2. Research and Breeding Institute of Pomology, Holovousy v Podkrkonoší, 508 01, Hořice, Czech Republic

    Hana Ludvíková & František Paprštein

  3. Plant Pathology, DipSA, Alma Mater Studiorum University of Bologna, viale Fanin 42, 40127, Bologna, Italy

    Assunta Bertaccini

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  1. Jana Fránová
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  2. Hana Ludvíková
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  3. František Paprštein
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  4. Assunta Bertaccini
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Correspondence to Jana Fránová.

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Fránová, J., Ludvíková, H., Paprštein, F. et al. Genetic diversity of Czech ‘Candidatus Phytoplasma mali’ strains based on multilocus gene analyses. Eur J Plant Pathol 136, 675–688 (2013). https://doi.org/10.1007/s10658-013-0196-5

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