Skip to main content
SpringerLink
Log in

The brome mosaic virus-based recombination vector triggers a limited gene silencing response depending on the orientation of the inserted sequence

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

In some RNA viruses (e.g. in brome mosaic virus, BMV), the same factor (intra- or intermolecular hybridization between viral RNA molecules) is capable of inducing two different processes: RNA silencing and RNA recombination. To determine whether there is some interplay between these two phenomena, we have examined if the BMV-based recombination vector containing a plant-genome-derived sequence can function as a gene-silencing vector. Surprisingly, we found that neither dsRNA forming during the replication of the BMV-based vector nor highly structured regions of its genome were effective RNAi triggers. Only mutants carrying a sequence complementary to the target mRNA functioned as gene silencing vectors and were steadily maintained in the infected plant. The constructs containing a sense sequence or inverted repeats did not induce gene silencing but instead were eliminated from the plant cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Barciszewska-Pacak M, Grabowska B, Wojciechowicz J, Pacak A, Figlerowicz M (2004) Virus-induced RNA silencing in plants. In: Sohail M (ed) Gene silencing by RNA interference: technology and application. CRC Press, Boca Raton, pp 323–342

    Google Scholar 

  2. Helliwell C, Waterhouse P (2003) Constructs and methods for high-throughput gene silencing in plants. Methods 30:289–295

    Article  CAS  PubMed  Google Scholar 

  3. Klahre U, Meins FJ (2004) RNA silencing in plants—biolistic delivery of RNAi reagents. In: Sohail M (ed) Gene silencing by RNA interference: technology and application. CRC Press, Boca Raton, pp 343–356

    Google Scholar 

  4. Silhavy D (2004) Agro-infiltration: a versatile tool for RNAi studies in plants. In: Sohail M (ed) Gene silencing by RNA interference: technology and application. CRC Press, Boca Raton, pp 357–364

    Google Scholar 

  5. Hull R (2002) Replication of positive-sense single-stranded RNA viruses. Matthews’ plant virology, 4th edn. Academic Press, London, pp 304–312

    Google Scholar 

  6. Ahlquist P (2002) RNA-dependent RNA polymerases, viruses, and RNA silencing. Science 296:1270–1273

    Article  CAS  PubMed  Google Scholar 

  7. Ruiz MT, Voinnet O, Baulcombe DC (1998) Initiation and maintenance of virus-induced gene silencing. Plant Cell 10:937–946

    Article  CAS  PubMed  Google Scholar 

  8. Lacomme C, Hrubikova K, Hein I (2003) Enhancement of virus-induced gene silencing through viral-based production of inverted-repeats. Plant J 34:543–553

    Article  CAS  PubMed  Google Scholar 

  9. Molnar A, Csorba T, Lakatos L, Varallyay E, Lacomme C, Burgyan J (2005) Plant virus-derived small interfering RNAs originate predominantly from highly structured single-stranded viral RNAs. J Virol 79:7812–7818

    Article  CAS  PubMed  Google Scholar 

  10. Saedler R, Baldwin IT (2004) Virus-induced gene silencing of jasmonate-induced direct defences, nicotine and trypsin proteinase-inhibitors in Nicotiana attenuata. J Exp Bot 55:151–157

    Article  CAS  PubMed  Google Scholar 

  11. Szittya G, Molnar A, Silhavy D, Hornyik C, Burgyan J (2002) Short defective interfering RNAs of tombusviruses are not targeted but trigger post-transcriptional gene silencing against their helper virus. Plant Cell 14:359–372

    Article  CAS  PubMed  Google Scholar 

  12. Lu R, Martin-Hernandez AM, Peart JR, Malcuit I, Baulcombe DC (2003) Virus-induced gene silencing in plants. Methods 30:296–303

    Article  CAS  PubMed  Google Scholar 

  13. Angell SM, Baulcombe DC (1997) Consistent gene silencing in transgenic plants expressing a replicating potato virus X RNA. EMBO J 16:3675–3684

    Article  CAS  PubMed  Google Scholar 

  14. Thomas CL, Jones L, Baulcombe DC, Maule AJ (2001) Size constraints for targeting post-transcriptional gene silencing and for RNA-directed methylation in Nicotiana benthamiana using a potato virus X vector. Plant J 25:417–425

    Article  CAS  PubMed  Google Scholar 

  15. Ratcliff F, Martin-Hernandez AM, Baulcombe DC (2001) Technical Advance. Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J 25:237–245

    Article  CAS  PubMed  Google Scholar 

  16. Kumagai MH, Donson J, Della-Cioppa G, Harvey D, Hanley K, Grill LK (1995) Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc Natl Acad Sci USA 92:1679–1683

    Article  CAS  PubMed  Google Scholar 

  17. Holzberg S, Brosio P, Gross C, Pogue GP (2002) Barley stripe mosaic virus-induced gene silencing in a monocot plant. Plant J 30:315–327

    Article  CAS  PubMed  Google Scholar 

  18. Hein I, Barciszewska-Pacak M, Hrubikova K, Williamson S, Dinesen M, Soenderby IE, Sundar S, Jarmołowski A, Shirasu K, Lacomme C (2005) Virus-induced gene silencing-based functional characterization of genes associated with powdery mildew resistance in barley. Plant Physiol 138:2155–2164

    Article  CAS  PubMed  Google Scholar 

  19. Ding XS, Schneider WL, Chaluvadi SR, Mian MA, Nelson RS (2006) Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Mol Plant Microbe Interact 19:1229–1239

    Article  CAS  PubMed  Google Scholar 

  20. Ahlquist P (1992) Bromovirus RNA replication and transcription. Curr Opin Genet Dev 2:71–76

    Article  CAS  PubMed  Google Scholar 

  21. Nagy PD, Bujarski JJ (1993) Targeting the site of RNA-RNA recombination in brome mosaic virus with antisense sequences. Proc Natl Acad Sci USA 90:6390–6394

    Article  CAS  PubMed  Google Scholar 

  22. Alejska M, Figlerowicz M, Malinowska N, Urbanowicz A, Figlerowicz M (2005) A universal BMV-based RNA recombination system-how to search for general rules in RNA recombination. Nucleic Acids Res 33:e105

    Article  PubMed  Google Scholar 

  23. Alejska M, Malinowska N, Urbanowicz A, Figlerowicz M (2005) Two types of non-homologous RNA recombination in brome mosaic virus. Acta Biochim Pol 52:833–844

    CAS  PubMed  Google Scholar 

  24. Matthews PD, Luo R, Wurtzel ET (2003) Maize phytoene desaturase and zeta-carotene desaturase catalyse a poly-Z desaturation pathway: implications for genetic engineering of carotenoid content among cereal crops. J Exp Bot 54:2215–2230

    Article  CAS  PubMed  Google Scholar 

  25. Voinnet O (2001) RNA silencing as a plant immune system against viruses. Trends Genet 17:449–459

    Article  CAS  PubMed  Google Scholar 

  26. Waterhouse PM, Wang MB, Lough T (2001) Gene silencing as an adaptive defence against viruses. Nature 411:834–842

    Article  CAS  PubMed  Google Scholar 

  27. Peele C, Jordan CV, Muangsan N, Turnage M, Egelkrout E, Eagle P, Hanley-Bowdoin L, Robertson D (2001) Silencing of a meristematic gene using geminivirus-derived vectors. Plant J 27:357–366

    Article  CAS  PubMed  Google Scholar 

  28. Figlerowicz M (2000) Role of RNA structure in non-homologous recombination between genomic molecules of brome mosaic virus. Nucleic Acids Res 28:1714–1723

    Article  CAS  PubMed  Google Scholar 

  29. Kalantidis K, Tsagris M, Tabler M (2006) Spontaneous short-range silencing of a GFP transgene in Nicotiana benthamiana is possibly mediated by small quantities of siRNA that do not trigger systemic silencing. Plant J 45:1006–1016

    Article  CAS  PubMed  Google Scholar 

  30. Schwartz M, Chen J, Janda M, Sullivan M, den Boon J, Ahlquist P (2002) A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids. Mol Cell 9:505–514

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the Polish Government through grant 6PO5A 054 21 from the State Committee for Scientific Research (KBN) awarded to MF grant PBZ-MNiSW-2/3/2006 to AP, AJ and ZSK. MF was additionally funded by grant CRP/POL03-02 from the International Center for Genetic Engineering and Biotechnology, and MBP by a Marie Curie Training PhD Fellowship Site in Plant Virology to the Scottish Crop Research Institute (contract number QLK3-CT-2001-60032). AP has been supported by the Foundation for Polish Science.

Author information

Author notes

  1. Christophe Lacomme

    Present address: Institute of Molecular Plant Sciences, University of Edinburgh, King’s Buildings, Mayfield Road, Edinburgh, EH9 3JH, UK

Authors and Affiliations

  1. Department of Gene Expression, Adam Mickiewicz University, Międzychodzka 5, 60-371, Poznan, Poland

    Andrzej Pacak, Maria Barciszewska-Pacak, Artur Jarmołowski & Zofia Szweykowska-Kulińska

  2. Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704, Poznan, Poland

    Pawel M. Strozycki, Magdalena Alejska & Marek Figlerowicz

  3. Plant Pathology Programme, Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK

    Christophe Lacomme

Authors
  1. Andrzej Pacak
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pawel M. Strozycki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maria Barciszewska-Pacak
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Magdalena Alejska
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christophe Lacomme
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Artur Jarmołowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zofia Szweykowska-Kulińska
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Marek Figlerowicz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Marek Figlerowicz.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pacak, A., Strozycki, P.M., Barciszewska-Pacak, M. et al. The brome mosaic virus-based recombination vector triggers a limited gene silencing response depending on the orientation of the inserted sequence. Arch Virol 155, 169–179 (2010). https://doi.org/10.1007/s00705-009-0556-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-009-0556-9

Keywords

Search

Navigation