Skip to main content

Advertisement

Springer Nature Link
Log in

The Citrus yellow mosaic badnavirus ORFI functions as a RNA-silencing suppressor

  • Short Report
  • Published:
Virus Genes Aims and scope Submit manuscript

Abstract

Citrus yellow mosaic badnavirus (CMBV) causes mosaic disease in all economically important citrus cultivars of India, with losses reaching up to 70%. CMBV belongs to the genus Badnavirus, family Caulimoviridae, possessing a circular double-stranded (ds) DNA genome with six open reading frames (ORFs I to VI), whose functions are yet to be deciphered. The RNA-silencing suppressor (RSS) activity has not been assigned to any CMBV ORF as yet. In the present study, it was found that ORFI exhibited RSS activity among all the six CMBV ORFs tested. Studies were done by employing the well-established Agrobacterium-mediated transient assay based on the transgenic Nicotiana benthamiana 16c plant line expressing the green fluorescent protein (GFP). The RSS activity of ORFI was confirmed by the analysis of the GFP visual expression in the agroinfiltrated leaves, further supported by quantification of GFP expression by RT-PCR. Based on the GFP visual expression, the CMBV ORFI was a weak RSS when compared to the p19 protein of tomato bushy stunt virus. In contrast, the ORFII, ORFIV, ORFV, ORFVI, and CP gene did not exhibit any RSS activity. Hence, ORFI is the first ORF of CMBV to be identified with RNA-silencing suppression activity.

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

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  1. Dakshinamurti V, Reddy GS (1975) Mosaic: a transmissible disorder of sweet oranges. Ind Phytopathol 28:398–399

    Google Scholar 

  2. Ahlawat YS, Chenulu VV, Vishwanath SM, Pandey PK (1984) Studies on a mosaic disease of citrus in India. Curr Sci 54:873–874

    Google Scholar 

  3. Ahlawat YS, Pant RP, Lockhart BE, Srivastava M, Chakraborty NK, Varma A (1996) Association of a badnavirus with citrus mosaic disease in India. Plant Dis 80:590–592

    Article  Google Scholar 

  4. Huang Q, Hartung S (2001) Cloning and sequence analysis of an infectious clone of Citrus yellow mosaic virus that can infect sweet orange via Agrobacterium-mediated inoculation. J Gen Virol 82:2549–2558

    Article  CAS  Google Scholar 

  5. Ahlawat YS, Varma A, Pant RP, Shukla A, Lockhart BEL (1996) Partial characterization of a badnavirus associated with citrus yellow mosaic disease in India. In: 13th Conference of the International Organization of Citrus Virologists. California: University of California Press, pp 208–217

  6. Anthony Johnson AM, Sai Gopal DVR, Sudhakar C, Dasgupta I (2017) Citrus yellow mosaic badnavirus infecting Citrus sp.: a threat to the citrus industry and a quarantine issue. J Gen Plant Pathol 83:57–65

    Article  CAS  Google Scholar 

  7. Anthony Johnson AM, Borah BK, Sai Gopal DVR, Dasgupta I (2012) Analysis of full-length sequences of two Citrus yellow mosaic badnavirus isolates infecting Citrus jambhiri (Rough Lemon) and Citrus sinensis L. Osbeck (Sweet Orange) from nursery in India. Virus Genes 45:600–605

    Article  CAS  Google Scholar 

  8. Borah BK, Sharma S, Kant R, Johnson AMA, Saigopal DVR, Dasgupta I (2013) Tropical badnaviruses and tungroviruses. Mol Plant Pathol 14:759–771

    Article  CAS  Google Scholar 

  9. Baker R, Caffier D, Choiseul JW, De Clercq P, Dormannsné-Simon E, Gerowitt B, Karadjova OE, Lövei G, Lansink AO, Makowski D, Manceau C, Manici L, Perdikis D, Puglia AP, Schans J, Schrader G, Steffek R, Strömberg A, Tiilikkala K, van Lenteren JC, Vloutoglou I (2008) Pest risk assessment made by France on Citrus yellow mosaic virus or Citrus mosaic badnavirus considered by France as harmful in the French overseas departments of French Guiana, Guadeloupe, Martinique and Réunion—Scientific Opinion of the Panel on Plant Health. EFSA J 6(686):1–16

    Google Scholar 

  10. Calil IP, Fontes EPB (2017) Plant immunity against viruses: antiviral immune receptors in focus. Ann Bot 119:711–723

    CAS  PubMed  Google Scholar 

  11. Rosa C, Kuo YW, Wuriyanghan H, Falk BW (2018) RNA interference mechanisms and applications in plant pathology. Annu Rev Phytopathol 56:581–610

    Article  CAS  Google Scholar 

  12. Voloudakis AE, Holeva MC, Sarin LP, Bamford DH, Vargas M, Poranen MM, Tenllado F (2015) Efficient double-stranded RNA production methods for utilization in plant virus control. Methods Mol Biol 1236:255–274

    Article  CAS  Google Scholar 

  13. Voloudakis A, Holeva M, Kaldis A, Kim D (2018) Tools and techniques for production of double stranded RNA and its application for management of plant viral diseases. In: Patil BL (ed) Genes, genetics and transgenics for virus resistance in plants. Caister Academic Press, Norfolk, pp 119-140. https://doi.org/10.21775/9781910190814

    Chapter  Google Scholar 

  14. Kaldis A, Berbati M, Melita O, Reppa C, Holeva M, Otten P, Voloudakis A (2018) Exogenously applied dsRNA molecules deriving from Zucchini yellow mosaic virus genome move systemically and protect cucurbits against ZYMV. Mol Plant Pathol 19:883–895

    Article  CAS  Google Scholar 

  15. Konakalla NC, Kaldis A, Berbati M, Masarapu H, Voloudakis AE (2016) Exogenous application of double-stranded RNA molecules from TMV p126 and CP genes confers resistance against TMV in tobacco. Planta 244:961–969

    Article  CAS  Google Scholar 

  16. Tharanath V, Patil BL, Kaldis A, Sai Gopal DVR, Ritesh M, Berbati M, Voloudakis A (2020) DsRNA-mediated protection against two isolates of Papaya ringspot virus through topical application of dsRNA in papaya. J Virol Methods 275:1–7

    Google Scholar 

  17. Voinnet O, Pinto YM, Baulcombe DC (1999) Suppression of gene silencing: a general strategy used by diverse DNA and RNA viruses of plants. Proc Natl Acad Sci USA 96:14147–14152

    Article  CAS  Google Scholar 

  18. Burgyán J, Havelda Z (2011) Viral suppressors of RNA silencing. Trends Plant Sci 16:265–272

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  20. Bendahmane A, Farnham G, Moffett P, Baulcombe DC (2002) Constitutive gain-of-function mutants in a nucleotide binding site-leucine rich repeat protein encoded at the Rx locus of potato. Plant J 32:195–204

    Article  CAS  Google Scholar 

  21. Vives MC, Martın S, Ambros S, Renovell A, Navarro L, Pina JA, Moreno P, Guerri J (2008) Development of a full-genome cDNA clone of Citrus leaf blotch virus and infection of citrus plants. Mol Plant Pathol 9:787–797

    Article  CAS  Google Scholar 

  22. Patil BL, Fauquet CM (2015) Light intensity and temperature affect systemic spread of silencing signal in transient agroinfiltration studies. Mol Plant Pathol 16:484–494

    Article  CAS  Google Scholar 

  23. Matsuo K, Fukuzawa N, Matsumura T (2016) A simple agroinfiltration method for transient gene expression in plant leaf discs. J Biosci Bioeng 122:351–356

    Article  CAS  Google Scholar 

  24. Mann KS, Dietzgen RG (2017) Functional analysis of a weak viral RNA silencing suppressor using two GFP variants as silencing inducers. J Virol Methods 239:50–57

    Article  CAS  Google Scholar 

  25. Vance V, Vaucheret H (2001) RNA silencing in plants–defense and counter defense. Science 292:2277–2280

    Article  CAS  Google Scholar 

  26. Csorba T, Kontra L, Burgyán J (2015) Viral silencing suppressors: tools forged to fine-tune host-pathogen coexistence. Virology 479–480:85–103

    Article  Google Scholar 

  27. Rajeswaran R, Golyaev V, Seguin J, Zvereva AS, Farinelli L, Pooggin MM (2014) Interactions of Rice tungro bacilliform pararetrovirus and its protein P4 with plant RNA-silencing machinery. Mol Plant Microbe Interact 27:1370–1378

    Article  Google Scholar 

  28. Andrew JL, Janet L, Justin H, Andrew JH, Ari S, Joel J (2007) Cauliflower mosaic virus protein P6 is a suppressor of RNA silencing. J Gen Virol 88:3439–3444

    Article  Google Scholar 

Download references

Acknowledgements

TV gratefully acknowledges scholarship from EACEA (Erasmus Mundus BRAVE program; Grant 2013-2536). DVRSG and BLP acknowledge visiting faculty scholarships from Erasmus Mundus BRAVE program. Authors are also grateful to Dr V. Pantaleo (CNR, IT) and Prof. O. Voinnet (ETH, CH) for providing the plasmids and A. tumefaciens strain GV3101 used in the present study. Dr Maria Holeva (BPI, GR) is acknowledged for providing the electroporator.

Author information

Authors and Affiliations

  1. Laboratory of Plant Breeding and Biometry, Faculty of Crop Science, Agricultural University of Athens, 11855, Athens, Greece

    Tharanath Vadlamudi, Athanasios Kaldis & Andreas E. Voloudakis

  2. Department of Virology, Sri Venkateswara University, Tirupati, Andhra Pradesh, 517502, India

    Tharanath Vadlamudi & Venkataramana Sai Gopal Divi

  3. ICAR-Indian Institute of Horticultural Research, Bengaluru, Karnataka, 560089, India

    Basavaprabhu L. Patil

Authors
  1. Tharanath Vadlamudi
    View author publications

    Search author on:PubMed Google Scholar

  2. Athanasios Kaldis
    View author publications

    Search author on:PubMed Google Scholar

  3. Venkataramana Sai Gopal Divi
    View author publications

    Search author on:PubMed Google Scholar

  4. Basavaprabhu L. Patil
    View author publications

    Search author on:PubMed Google Scholar

  5. Andreas E. Voloudakis
    View author publications

    Search author on:PubMed Google Scholar

Contributions

AV and DVRSG conceived the study; AV designed, supervised the study, and corrected the manuscript; TV performed the experiments and drafted the manuscript; AK assisted in the experimentation, data analysis, and manuscript writing; BLP assisted in experimental design and manuscript writing.

Corresponding author

Correspondence to Andreas E. Voloudakis.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human and animal rights

This research did not involve any animal and/or human participants.

Additional information

Edited by Karel Petrzik.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Vadlamudi, T., Kaldis, A., Divi, V.S.G. et al. The Citrus yellow mosaic badnavirus ORFI functions as a RNA-silencing suppressor. Virus Genes 57, 469–473 (2021). https://doi.org/10.1007/s11262-021-01863-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11262-021-01863-8

Keywords

Profiles

  1. Basavaprabhu L. Patil View author profile
  2. Andreas E. Voloudakis View author profile