Biopolym. Cell. 2014; 30(2):141-148.
Viruses and Cell
Nucleotide and amino acid sequences of a coat protein of an Ukrainian isolate of Potato virus Y: comparison with homologous sequences of other isolates and phylogenetic analysis
1Budzanivska I. G., 2Ovcharenko L. P., 1Kharina A. V., 3, 4Boubriak I. I., 1Polischuk V. P.
  1. Educational and Scientific Center "Institute of Biology",
    Taras Shevchenko National University of Kyiv
    64/13, Volodymyrska Str., Kyiv, Ukraine, 01601
  2. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  3. Department of Biochemistry, University of Oxford
    South Parks Road, Oxford, OX1 3QU, UK
  4. Institute of Cell Biology and Genetic Engineering, NAS of Ukraine
    148, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680

Abstract

Aim. Identification of the widespread Ukrainian isolate(s) of PVY (Potato virus Y) in different potato cultivars and subsequent phylogenetic analysis of detected PVY isolates based on NA and AA sequences of coat protein. Methods. ELISA, RT-PCR, DNA sequencing and phylogenetic analysis. Results. PVY has been identified serologically in potato cultivars of Ukrainian selection. In this work we have optimized a method for total RNA extraction from potato samples and offered a sensitive and specific PCR-based test system of own design for diagnostics of the Ukrainian PVY isolates. Part of the CP gene of the Ukrainian PVY isolate has been sequenced and analyzed phylogenetically. It is demonstrated that the Ukrainian isolate of Potato virus Y (CP gene) has a higher percentage of homology with the recombinant isolates (strains) of this pathogen (approx. 98.8– 99.8 % of homology for both nucleotide and translated amino acid sequences of the CP gene). The Ukrainian isolate of PVY is positioned in the separate cluster together with the isolates found in Syria, Japan and Iran; these isolates possibly have common origin. The Ukrainian PVY isolate is confirmed to be recombinant. Conclusions. This work underlines the need and provides the means for accurate monitoring of Potato virus Y in the agroecosystems of Ukraine. Most importantly, the phylogenetic analysis demonstrated the recombinant nature of this PVY isolate which has been attributed to the strain group O, subclade N:O.
Keywords: Potato virus Y, potyvirus, PCR, phylogenetic analysis, recombinant strain

References

[1] Moury B, Morel C, Johansen E, Jacquemond M. Evidence for diversifying selection in Potato virus Y and in the coat protein of other potyviruses. J Gen Virol. 2002; 83(Pt 10):2563–73.
[2] Singh RP, Valkonen JP, Gray SM, Boonham N, Jones RA, Kerlan C, Schubert J. Discussion paper: the naming of Potato virus Y strains infecting potato. Arch Virol. 2008; 153(1):1–13.
[3] Valkonen JPT. Potato viruses: economical losses and biotechnological potential. Potato biology and biotechnology. San Diego: Elsevier, 2007; 619–641.
[4] Virus taxonomy. Ninth report of the International Committee on Taxonomy of Viruses / Eds AMQ King, E Lefkowitz, MJ Adams, EB Carstens. Wien: Springer, 2012. 1327 p.
[5] Smith KM. Composite nature of certain potato viruses of the mosaic group. Nature. 1931; 127:702.
[6] Bawden FC, Kassanis B. Varietal differences in susceptibility to potato virus Y. Ann Appl Biol. 1946; 33(1):46–50.
[7] Cockerham G. The reactions of the potato varieties to viruses X, A, B, and C. Ann Appl Biol. 1943; 30(4):338–344.
[8] Kus M. The epidemic of the tuber necrotic ringspot strain of potato virus Y (PVYNTN) and its effect on potato crops in Slovenia. 9th EAPR Virology Section Meeting Bled (18–22 June, 1995): 159–160.
[9] Kerlan C, Le Romancer M. Potato tuber necrotic ringspot disease. Proc. EAPR Meeting, Virology section. Vitoria-Gasteiz (Spain), 1992:77–79.
[10] Roossinck MJ. Mechanisms of plant virus evolution. Annu Rev Phytopathol. 1997; 35:191–209.
[11] Schubert J, Fomitcheva V, Sztangret-Wisniewska J. Differentiation of Potato virus Y strains using improved sets of diagnostic PCR-primers. J Virol Methods. 2007; 140(1–2):66–74.
[12] Nie X, Singh RP. Specific differentiation of recombinant PVYN: O and PVYNTN isolates by multiplex RT-PCR. J Virol Methods. 2003; 113(2):69–77.
[13] Nie X, Singh RP, Singh M. Molecular and pathological characterization of N:O isolates of the Potato virus Y from Manitoba, Canada. Can J Plant Pathol. 2004; 26(4):573–83.
[14] Lakin GF. Biometrics: studies: manual [for biological. specials. universities]. M.: Higher School, 1980; 293 p.
[15] Dzheyper J, Scott RF. Genetic engineering of plants. Moscow: Mir, 1991; 236–269.
[16] Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011; 28(10):2731–9.
[17] Felsenstein J. Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol. 1981; 17(6):368–376.
[18] Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987; 4(4): 406–425.
[19] Huelsenbeck JP, Crandall KA. Phylogeny estimation and hypothesis testing using maximum likelihood. Annu Rev Ecol Syst. 1997; 28:437–66.
[20] Chikh Ali M, Maoka T, Natsuaki KT. The occurrence and characterization of new recombinant Isolates of PVY displaying shared properties of PVYNW and PVYNTN. J Phytopathol. 2007; 155 (7–8):409