Archives of Virology

, Volume 159, Issue 6, pp 1373–1383 | Cite as

Sequence variability in the HC-Pro coding regions of Korean soybean mosaic virus isolates is associated with differences in RNA silencing suppression

  • Mei-Jia Li
  • Jung-Kyu Kim
  • Eun-Young Seo
  • Seok Myeong Hong
  • Eui-Il Hwang
  • Jung-Kyung Moon
  • Leslie L. Domier
  • John HammondEmail author
  • Young-Nam YounEmail author
  • Hyoun-Sub LimEmail author
Original Article


Soybean mosaic virus (SMV), a member of the family Potyviridae, is an important viral pathogen affecting soybean production in Korea. Variations in helper component proteinase (HC-Pro) sequences and the pathogenicity of SMV samples from seven Korean provinces were compared with those of previously characterized SMV isolates from China, Korea and the United States. Phylogenetic analysis separated 16 new Korean SMV isolates into two groups. Fourteen of the new Korean SMV samples belonged to group II and were very similar to U.S. strain SMV G7 and Chinese isolate C14. One isolate in group II, A297-13, differed at three amino acid positions (L54F, N286D, D369N) in the HC-Pro coding sequence from severe isolates and SMV 413, showed very weak silencing suppressor activity, and produced only mild symptoms in soybean. To test the role of each amino acid substitution in RNA silencing and viral RNA accumulation, a series of point mutations was constructed. Substitution of N for D at position 286 in HC-Pro of SMV A297-12 significantly reduced silencing suppression activity. When the mutant HC-Pro of A297-13 was introduced into an infectious clone of SMV 413, accumulation of viral RNA was reduced to less than 3 % of the level of SMV 413 containing HC-Pro of A297-12 at 10 days post-inoculation (dpi) but increased to 40 % of SMV 413(HC-Pro A297-12) at 40 dpi. At 50 dpi RNA accumulation of SMV 413(HC-Pro A297-13) was similar to that of SMV 413(HC-Pro A297-12). However, at 50 dpi, the D at position 286 of HC-Pro in SMV 413(HC-Pro A297-13) was found to have reverted to N. The results showed that 1) a naturally occurring mutation in HC-Pro significantly reduced silencing suppression activity and accumulation of transgene and viral RNAs, and 2) that there was strong selection for revision to wild type when the mutation was introduced into an infectious clone of SMV.


Tobacco Etch Virus Soybean Mosaic Virus Zucchini Yellow Mosaic Virus Silence Suppressor Korean Isolate 
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.



This research was supported by iPET (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries), Ministry of Agriculture, Food and Rural Affairs (Project No. 112018-3).

Supplementary material

705_2013_1964_MOESM1_ESM.docx (13.9 mb)
Supplementary material 1 (DOCX 14196 kb)


  1. 1.
    Anjos JR, Jarlfors U, Ghabrial SA (1992) Soybean mosaic potyvirus enhances the titer of two comoviruses in dually infected soybean plants. Phytopathology 82:1022–1027CrossRefGoogle Scholar
  2. 2.
    Arguello-Astorga G, Ascencio-Ibáñez JT, Dallas MB, Orozco BM, Hanley-Bowdoin L (2007) High-frequency reversion of geminivirus replication protein mutants during infection. J Virol 81:11005–11015PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Atreya CD, Pirone TP (1993) Mutational analysis of the helper component-proteinase gene of a potyvirus: effects of amino acid substitutions, deletions, and gene replacement on virulence and aphid transmissibility. Proc Natl Acad Sci USA 90:11919–11923PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Atreya CD, Raccah B, Pirone TP (1990) A point mutation in the coat protein abolish aphid transmissibility of a potyvirus. Virology 178(1):161–165PubMedCrossRefGoogle Scholar
  5. 5.
    Atsumi G, Nakahara KS, Wada TS, Choi SH, Masuta C, Uyeda I (2012) Heterologous expression of viral suppressors of RNA silencing complements virulence of the HC-Pro mutant of clover yellow vein virus in pea. Arch Virol 157:1019–1028PubMedCrossRefGoogle Scholar
  6. 6.
    Bayne EH, Rakitina DV, Morozov SY, Baulcombe DC (2005) Cell-to-cell movement of potato potexvirus X is dependent on suppression of RNA silencing. Plant J 44:471–482PubMedCrossRefGoogle Scholar
  7. 7.
    Blanc S, Lopez-Moya JJ, Wang R, Garcia-Lampasona S, Thornbury DW, Pirone TP (1997) A specific interaction between coat protein and helper component correlates with aphid transmission of a potyvirus. Virology 231:141–147PubMedCrossRefGoogle Scholar
  8. 8.
    Blanc S, Ammar ED, Garcia-Lampasona S, Dolja VV, Llave C, Baker J, Pirone TP (1998) Mutations in the potyvirus helper component protein: effects on interactions with virions and aphid stylets. J Gen Virol 79:3119–3122PubMedGoogle Scholar
  9. 9.
    Bragg JN, Jackson AO (2004) The C-terminal region of the Barley stripe mosaic virus γb protein participates in homologous interactions and is required for suppression of RNA silencing. Mol Plant Pathol 5:465–481PubMedCrossRefGoogle Scholar
  10. 10.
    Bull JJ, Meyers LA, Lachmann M (2005) Quasispecies made simple. PLoS Comput Biol 1(6):e61PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Chakrabarty R, Banerjee R, Chung SM, Farman M, Citovsky V, Hogenhout SA, Tzfira T, Goodin M (2007) pSITE vectors for stable integration or transient expression of autofluorescent protein fusions in plants: probing Nicotiana benthamiana–virus interactions. Mol Plant Microbe Interact 20:740–750PubMedCrossRefGoogle Scholar
  12. 12.
    Cho EK, Choi SH, Cho WT (1983) Newly recognized soybean mosaic virus mutants and sources of resistance in soybeans. Res Rep ORD (SPMU) 25:18–22Google Scholar
  13. 13.
    Cho EK, Chung BJ (1976) Studies on identification and classification of soybean virus diseases I. Preliminary studies on a soybean virus disease. Korean J Plant Prot 15:61–68Google Scholar
  14. 14.
    Cho EK, Chung BJ, Lee SH (1977) Studies on identification and classification of soybean virus diseases in Korea. Etiology of a necrotic disease of Glycine max. Plant Dis Rep 61:313–317Google Scholar
  15. 15.
    Cho EK, Goodman RM (1979) Strains of soybean mosaic virus: classification based on virulence in resistant soybean cultivars. Phytopathology 69:467–470CrossRefGoogle Scholar
  16. 16.
    Cho EK, Goodman RM (1982) Evaluation of resistance in soybeans to soybean mosaic virus strains. Crop Sci 22:1133–1136CrossRefGoogle Scholar
  17. 17.
    Chung BJ, Lee SH, Cho EK, Park HC (1974) Identification of soybean viruses and soybean varietal reactions. Annu Rep Circ 26:137–145Google Scholar
  18. 18.
    Clarke DK, Duarte EA, Elena SF, Moya A, Dominigo E, Holland J (1994) The red queen reigns in the kingdom of RNA viruses. Proc Natl Acad Sci USA 91:4821–4824PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Deng M, Bragg JN, Ruzin S, Schichnes D, King D, Goodin MM, Jackson AO (2007) Role of the sonchus yellow net virus N protein in formation of nuclear viroplasms. J Virol 81:5362–5374PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Desbiez C, Girard M, Lecoq H (2010) A novel natural mutation in HC-Pro responsible for mild symptomatology of Zucchini yellow mosaic virus (ZYMV, Potyvirus) in cucurbits. Arch Virol 155:397–401PubMedCrossRefGoogle Scholar
  21. 21.
    Dolja VV, Herndon KL, Pirone TP, Carrington JC (1993) Spontaneous mutagenesis of a plant potyvirus genome after insertion of a foreign gene. J Virol 67:5968–5975PubMedCentralPubMedGoogle Scholar
  22. 22.
    Dombrovsky A, Sobolev I, Chejanovsky N, Raccah B (2007) Characterization of RR-1 and RR-2 cuticular proteins from Myzus persicae. Comp Biochem Physiol B 146:256–264PubMedCrossRefGoogle Scholar
  23. 23.
    Domier LL, Hobbs HA, McCoppin NK, Bowen CR, Steinlage TA, Chang S, Wang Y, Hartman GL (2011) Multiple loci condition seed transmission of Soybean mosaic virus (SMV) and SMV-induced seed coat mottling in soybean. Phytopathology 101:750–756PubMedCrossRefGoogle Scholar
  24. 24.
    Domier LL, Latorre IJ, Steinlage TA, McCoppin N, Hartman GL (2003) Variability and transmission by Aphis glycines of North American and Asian Soybean mosaic virus isolates. Arch Virol 148:1925–1941PubMedCrossRefGoogle Scholar
  25. 25.
    Drake JW, Holland JJ (1999) Mutation rates among RNA viruses. Proc Natl Acad Sci USA 96:13910–13913PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    English JJ, Davenport GF, Elmayan T, Vaucheret H, Baulcombe DC (1997) Requirement of sense transcription for homology-dependent virus resistance and trans-inactivation. Plant J 12:597–603CrossRefGoogle Scholar
  27. 27.
    Garcia-Arenal F, Fraile A, Malpica JM (2003) Variation and evolution of plant virus populations. Int Microbiol 6:225–232PubMedCrossRefGoogle Scholar
  28. 28.
    Govier DA, Kassanis B (1974) Evidence that a component other than the virus particle is needed for aphid transmission of potato virus Y. Virology 57:285–286PubMedCrossRefGoogle Scholar
  29. 29.
    Hajimorad MR, Eggenberger AL, Hill JH (2008) Adaptation of soybean mosaic virus avirulent chimeras containing P3 sequences from virulent strains to Rsv1-genotype soybeans is mediated by mutations in HC-Pro. Mol Plant Microbe Interact 21:937–946PubMedCrossRefGoogle Scholar
  30. 30.
    Huet H, Gal-On A, Meir E, Lecoq H, Raccah B (1994) Mutations in the helper component protease gene of zucchini yellow mosaic virus affect its ability to mediate aphid transmissibility. J Gen Virol 75:1407–1414PubMedCrossRefGoogle Scholar
  31. 31.
    Johansen LK, Carrington JC (2001) Silencing on the spot induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system. Plant Physiol 126:930–938PubMedCentralPubMedCrossRefGoogle Scholar
  32. 32.
    Johansen IE, Dougherty WG, Keller KE, Wang D, Hampton RO (1996) Multiple viral determinants affect seed transmission of pea seedborne mosaic virus in Pisum sativum. J Gen Virol 77:3149–3154PubMedCrossRefGoogle Scholar
  33. 33.
    Jossey S, Hobbs HA, Domier LL (2013) Role of soybean mosaic virus-encoded proteins in seed and aphid transmission in soybean. Phytopathology 103:941–948PubMedCrossRefGoogle Scholar
  34. 34.
    Kawakami S, Hori K, Hosokawa D, Okada Y, Watanabe Y (2003) Defective tobamovirus movement protein lacking wild-type phosphorylation sites can be complemented by substitutions found in revertants. J Virol 77:1452–1461PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Kim YH (2000) G7H, a new strain of soybean mosaic virus: its virulence and nucleotide sequence of cylindrical inclusion gene. Ph.D. thesis, Kyungpook National University, Daegu, p 120Google Scholar
  36. 36.
    Kim YH, Kim OS, Lee BC, Moon JK, Lee SC, Lee JY (2003) G7H, a new soybean mosaic virus strain: its virulence and nucleotide sequence of CI gene. Plant Dis 87:1372–1375CrossRefGoogle Scholar
  37. 37.
    Lim SM (1985) Resistance to soybean mosaic virus in soybeans. Phytopathology 75:199–201CrossRefGoogle Scholar
  38. 38.
    Lim HS, Jang C, Bae H, Kim J, Lee C, Hong J, Ju H, Kim H, Domier LL (2011) Soybean mosaic virus infection and helper component-protease enhance accumulation of bean pod mottle virus-specific siRNAs. Plant Pathol J 27:315–323CrossRefGoogle Scholar
  39. 39.
    Lin SS, Wu HW, Jan FJ, Hou RF, Yeh SD (2007) Modifications of the helper component-protease of Zucchini yellow mosaic virus for generation of attenuated mutants for cross protection against severe infection. Phytopathology 97:287–296PubMedCrossRefGoogle Scholar
  40. 40.
    Malpica JM, Fraile A, Moreno I, Obies CI, Drake JW, Garcia-Arenal F (2002) The rate and character of spontaneous mutation in an RNA virus. Genetics 162:1505–1511PubMedCentralPubMedGoogle Scholar
  41. 41.
    Moya A, Elena SF, Bracho A, Miralles R, Barrio E (2000) The evolution of RNA viruses: a population genetics view. Proc Natl Acad Sci USA 97:6967–6973PubMedCentralPubMedCrossRefGoogle Scholar
  42. 42.
    Peng YH, Kadoury D, Gal-On A, Huet H, Wang Y, Raccah B (1998) Mutations in the HC-Pro gene of zucchini yellow mosaic potyvirus: effects on aphid transmission and binding to purified virions. J Gen Virol 79:897–904PubMedGoogle Scholar
  43. 43.
    Petty IT, Hunter BG, Wei N, Jackson AO (1989) Infectious barley stripe mosaic virus RNA transcribed in vitro from full-length genomic cDNA clones. Virology 171:342–349PubMedCrossRefGoogle Scholar
  44. 44.
    Pruss G, Ge X, Shi XM, Carrington JC, Vance VB (1997) Plant vira1 synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Plant Cell 9:859–868PubMedCentralPubMedCrossRefGoogle Scholar
  45. 45.
    Rajamäki M, Merits A, Rabenstein F, Andrejeva J, Paulin L, Kekarainen T, Kreuze JF, Forster RL, Valkonen JP (1998) Biological, serological, and molecular differences among isolates of potato a potyvirus. Phytopathology 88:311–321PubMedCrossRefGoogle Scholar
  46. 46.
    Rojas MR, Zerbini FM, Allison RF, Gilbertson RL, Lucas WJ (1997) Capsid protein and helper component-proteinase function as potyvirus cell-to-cell movement proteins. Virology 237:283–295PubMedCrossRefGoogle Scholar
  47. 47.
    Ruiz-Jarabo CM, Arias A, Baranowski E, Escarmis C, Domingo E (2000) Memory in viral quasispecies. J Virol 74(8):3543–3547PubMedCentralPubMedCrossRefGoogle Scholar
  48. 48.
    Shi XM, Miller H, Verchot J, Carrington JC, Vance VB (1997) Mutations in the region encoding the central domain of helper component-proteinase (HC-Pro) eliminate potato virus X/potyviral synergism. Virology 231:35–42PubMedCrossRefGoogle Scholar
  49. 49.
    Tamura K, Peterson D, Perterson 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 28:2731–2739PubMedCentralPubMedCrossRefGoogle Scholar
  50. 50.
    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. Nucleic Acid Res 25:4876–4882PubMedCentralPubMedCrossRefGoogle Scholar
  51. 51.
    Torres-Barceló C, Martín S, Darós JA, Elena SF (2008) From hypo- to hypersuppression: effect of amino acid substitutions on the RNA-silencing suppressor activity of the Tobacco etch potyvirus HC-Pro. Genetics 180:1039–1049PubMedCentralPubMedCrossRefGoogle Scholar
  52. 52.
    Vaira AM, Lim H-S, Bauchan GR, Owens RA, Natilla A, Dienelt MM, Reinsel MD, Hammond J (2012) Lolium latent virus (Alphaflexiviridae) coat proteins: expression and functions in infected plant tissue. J Gen Virol 93:1814–1824PubMedCrossRefGoogle Scholar
  53. 53.
    Varrelmann M, Maiss E, Pilot R, Palkovics L (2007) Use of pentapeptide-insertion scanning mutagenesis for functional mapping of the plum pox virus helper component proteinase suppressor of gene silencing. J Gen Virol 88:1005–1015PubMedCrossRefGoogle Scholar
  54. 54.
    Wu HW, Lin SS, Chen KC, Yeh SD, Chua NH (2010) Discriminating mutations of HC-Pro of Zucchini yellow mosaic virus with differential effects on small RNA pathways involved in viral pathogenicity and symptom development. Mol Plant Microbe Interact 23:17–28PubMedCrossRefGoogle Scholar
  55. 55.
    Wurch T, Lestienne F, Pauwels P (1998) A modified overlap extension PCR method to create chimeric genes in the absence of restriction enzymes. Biotechnol Tech 12:653–657CrossRefGoogle Scholar
  56. 56.
    Yambao MLM, Yagihashi H, Sekiguchi H, Sekiguchi T, Sasaki T, Sato M, Atsumi G, Tacahashi Y, Nakahara KS, Uyeda I (2008) Point mutations in helper component protease of clover yellow vein virus are associated with the attenuation of RNA-silencing suppression activity and symptom expression in broad bean. Arch Virol 153:105–115PubMedCrossRefGoogle Scholar
  57. 57.
    Yang S, Ravelonandro M (2002) Molecular studies of the synergistic interactions between plum pox virus HC-Pro protein and potato virus X. Arch Virol 147:2301–2312PubMedCrossRefGoogle Scholar
  58. 58.
    Xu Z, Polston JE, Goodman RM (1986) Identification of soybean mosaic, southern bean mosaic and tobacco ringspot viruses from soybean in the People’s Republic of China. Ann Appl Biol 108:51–57CrossRefGoogle Scholar
  59. 59.
    Zhang CQ, Ghabrial SA (2006) Development of Bean pod mottle virus-based vectors for stable protein expression and sequence-specific virus-induced gene silencing in soybean. Virology 344:401–411PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • Mei-Jia Li
    • 1
  • Jung-Kyu Kim
    • 1
  • Eun-Young Seo
    • 1
  • Seok Myeong Hong
    • 3
  • Eui-Il Hwang
    • 2
  • Jung-Kyung Moon
    • 4
  • Leslie L. Domier
    • 5
  • John Hammond
    • 6
    Email author
  • Young-Nam Youn
    • 1
    Email author
  • Hyoun-Sub Lim
    • 1
    Email author
  1. 1.Department of Applied BiologyChungnam National UniversityDaejeonKorea
  2. 2.KT&G R&D Headquarters Technical CenterDaejeonKorea
  3. 3.GenoRevo Korea Co.SuwonKorea
  4. 4.National Institute of Crop ScienceRural Development AdministrationSuwonKorea
  5. 5.Department of Crop SciencesUniversity of Illinois, USDA-ARSUrbanaUSA
  6. 6.Floral and Nursery Plants Research UnitUS National Arboretum, USDA-ARSBeltsvilleUSA

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