Archives of Virology

, Volume 159, Issue 9, pp 2457–2462 | Cite as

Molecular characterization of a Chinese isolate of potato virus A (PVA) and evidence of a genome recombination event between PVA variants at the 3′-proximal end of the genome

Brief Report

Abstract

Potato plants that exhibited mosaic symptoms were collected in Xiangxi, Hunan province, China. Multiplex RT-PCR screening for common viruses revealed the presence of potato virus A (PVA) in these samples. ELISA with virus-specific antibodies confirmed infection by PVA in the plants. Rod-shaped virions of ~750 nm in length and ~13 nm in width were observed by transmission electron microscopy. One virus isolate (designated PVA-Hunan) was subjected to molecular characterization. The viral genome consisted of 9,567 nucleotides, excluding the poly(A) tail, and encoded a polyprotein of 3,059 amino acids. A second characteristic potyvirus open reading frame (ORF), pretty interesting Potyviridae ORF (pipo), was located at nucleotides 2,834-3,139. The isolate shared 84 % to 98 % and 93 % to 99 % sequence identity with other PVA isolates at the nucleotide and amino acid level, respectively. Phylogenetic analysis demonstrated that, within the PVA group, PVA-Hunan clustered most closely with the Finnish isolate Her, then with isolates 143, U, Ali, M and B11. The isolate TamMV stood alone at a separate branch. However, scanning of complete genome sequences using SimPlot revealed 99 %-sequence identity between PVA-Hunan and TamMV in the 3′-proximal end of the genome (~nt 9,160 to the 3′end) and a 50 %-94 % (average ~83 %) identity upstream of nt 9,160. In contrast, 98 % identity between PVA-Hunan and isolates M and B11 was detected for nucleotides 1 to ~9,160, but only ~94 % for the 3′-proximal region, suggesting a genome recombination event (RE) at nt 9,133. The recombination breakpoint also was identified by the Recombination Detection Program (RDP). The RE was further confirmed by analysis of the CP gene, where the apparent RE was located.

Keywords

Potato Virus Mosaic Symptom Recombination Breakpoint Chinese Isolate Recombination Detection Program 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This research was supported by the National High Technology Research and Development Program (Project no. 2013AA102603), Hunan Provincial Natural Science Foundation (Project no. 13JJ2029), and the Ministry of Science and Technology of China through the ISTP Canada-China Science and Technology program (Project no. CCRD07-077). The authors thank Virginia Dickson for careful proofreading of the manuscript prior to submission.

Supplementary material

705_2014_2053_MOESM1_ESM.docx (28 kb)
Supplementary material 1 (DOCX 27 kb)

References

  1. 1.
    Bai Y, Nie X, Li X, Gao Y, Fan G, Zhang W, Geng H, Liu W (2010) The seed potato inspection and virus research in China. In: 93rd annual meeting of the potato association of America, The American Journal of Potato Research, vol 87. pp 112–123, Fredericton, 9–13 August 2009Google Scholar
  2. 2.
    Bartels R (1971) Potato virus A. Descriptions of plant viruses no. 54. CMI/AAB, UKGoogle Scholar
  3. 3.
    Chung BY, Miller WA, Atkins JF, Firth AE (2008) An overlapping essential gene in the Potyviridae. Proc Natl Acad Sci USA 105:5897–5902PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Dougherty WG, Carrington JC (1988) Expression and function of potyviral gene products. Annu Rev Phytopathol 26:123–143CrossRefGoogle Scholar
  5. 5.
    German TL (2001) Potato virus A. In: Stevenson WR, Loria R, Franc GD, Weingartner DP (eds) Compendium of potato diseases, 2nd edn. APS Press, St. Paul, pp 66–67Google Scholar
  6. 6.
    Guo D, Rajamaki ML, Saarma M, Valkonen JP (2001) Towards a protein interaction map of potyviruses: protein interaction matrixes of two potyviruses based on the yeast two-hybrid system. J Gen Virol 82:935–939PubMedGoogle Scholar
  7. 7.
    He C, Nie X (2012) Potato viruses and viral disease control. In: Tai GCC, Xiong X (eds) Research topics in world potato science. ISBN 978-0-9879812-0-2, OmniScience Press, Toronto, pp 189–214Google Scholar
  8. 8.
    Jansky SH, Jin LP, Xie KY, Xie CH, Spooner DM (2009) Potato production and breeding in China. Potato Res 52:57–65CrossRefGoogle Scholar
  9. 9.
    Kekarainen T, Merits A, Oruetxebarria I, Rajamäki ML, Valkonen JPT (1999) Comparison of the complete sequences of five different isolates of Potato virus A (PVA), genus Potyvirus. Arch Virol 144:2355–2366PubMedCrossRefGoogle Scholar
  10. 10.
    Kekarainen T, Savilahti H, Valkonen J (2002) Functional genomics on potato virus A: virus genome-wide map of sites essential for virus propagation. Genome Res 12:584–594PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Lole KS, Bollinger RC, Paranjape RS, Gadkari D, Kulkarni SS, Novak NG, Ingersoll R, Sheppard HW, Ray SC (1999) Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination. J Virol 73:152–160PubMedCentralPubMedGoogle Scholar
  12. 12.
    MacLachlan D, Larson R, Walker J (1954) Potato virus A. Am Potato J 31:67–72CrossRefGoogle Scholar
  13. 13.
    Martin DP, Lemey P, Lott M, Moulton V, Posada D, Lefeuvre P (2010) RDP3: a flexible and fast computer program for analyzing recombination. Bioinformatics 26:2462–2463PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Merits A, Rajamaki ML, Lindholm P, Runeberg-Roos P, Kekarainen T, Puustinen P, Makelainen K, Valkonen JP, Saarma M (2002) Proteolytic processing of potyviral proteins and polyprotein processing intermediates in insect and plant cells. J Gen Virol 83:1211–1221PubMedGoogle Scholar
  15. 15.
    Nie X, Singh RP (2001) Differential accumulation of potato virus A and expression of pathogenesis-related genes in resistant potato cv. Shepody upon graft inoculation. Phytopathology 91:197–203PubMedCrossRefGoogle Scholar
  16. 16.
    Nie X, Singh RP (2001) A novel usage of random primers for multiplex RT-PCR detection of virus and viroid in aphids, leaves, and tubers. J Virol Methods 91:37–49PubMedCrossRefGoogle Scholar
  17. 17.
    Puurand Ü, Mäkinen K, Baumann M, Saarma M (1992) Nucleotide sequence of the 3′-terminal region of potato virus A RNA. Virus Res 23:99–105PubMedCrossRefGoogle Scholar
  18. 18.
    Puurand U, Mäkinen K, Paulin L, Saarma M (1994) The nucleotide sequence of potato virus A genomic RNA and its sequence similarities with other potyviruses. J Gen Virol 75:457–461PubMedCrossRefGoogle Scholar
  19. 19.
    Puurand Ü, Valkonen J, Mäkinen K, Rabensteind F, Saarmaa M (1996) Infectious in vitro transcripts from cloned cDNA of the potato A potyvirus. Virus Res 40:135–140PubMedCrossRefGoogle Scholar
  20. 20.
    Rajamäki M, Merits A, Rabenstein F, Andrejeva J, Paulin L, Kekarainen T, Kreuze JF, Forster RLS, Valkonen JPT (1998) Biological, serological, and molecular differences among isolates of potato A potyvirus. Phytopathology 88:311–321PubMedCrossRefGoogle Scholar
  21. 21.
    Singh R, Nie X, Tai GCC (2000) A novel hypersensitive resistance response against potato virus A in cultivar ‘Shepody’. Theor Appl Genet 100:401–408CrossRefGoogle Scholar
  22. 22.
    Slack SA (2001) Potato virus X. In: Stevenson WR, Loria R, Franc GD, Weingartner DP (eds) Compendium of potato diseases, 2nd edn. APS Press, St. Paul, p 69Google Scholar
  23. 23.
    Slack SA, Singh RP (1998) Control of viruses affecting potatoes through seed potato certification programs. In: Hadidi A, Khetarpal RK, Koganezawa H (eds) Plant virus disease control. American Phytopathological Society Press, St. Paul, pp 249–260Google Scholar
  24. 24.
    Tamura K, Peterson D, Peterson N, Stecher G. Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739Google Scholar
  25. 25.
    Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acids Res 25:4876–4882PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Visser RG, Bachem CW, de Boer JM, Bryan GJ, Chakrabati SK, Feingold SR, Gromadka R, van Ham CHJ, Huang S, Jacobs JME, Kuznetsov B, de Melo PE, Milbourne D, Orjeda G, Sagredo B, Tang X (2009) Sequencing the potato genome: outline and first results to come from the elucidation of the sequence of the world’s third most important food crop. Am J Potato Res 86:417–429CrossRefGoogle Scholar
  27. 27.
    Wang B, Ma YL, Zhang ZB, Wu ZM, Wu YF, Li MF, Wang QC (2011) Potato viruses in China. Crop Prot 30:1117–1123CrossRefGoogle Scholar

Copyright information

© Her Majesty the Queen in Right of Canada 2014

Authors and Affiliations

  1. 1.Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Provincial Engineering Research Center for Potatoes, College of Horticulture and LandscapeHunan Agricultural UniversityChangshaChina
  2. 2.Agricultural Certification ServicesFrederictonCanada
  3. 3.Potato Research Centre, Agriculture and Agri-Food CanadaFrederictonCanada

Personalised recommendations