Advertisement

VirusDisease

, Volume 29, Issue 2, pp 207–211 | Cite as

RNA-based vaccination of Bhut Jolokia pepper (Capsicum chinense Jacq.) against cucumber mosaic virus

  • Munmi Borah
  • Margarita Berbati
  • Chrysavgi Reppa
  • Maria Holeva
  • Palash Deb Nath
  • Andreas Voloudakis
Short Communication

Abstract

Cucumber mosaic virus (CMV) causes great losses in Bhut Jolokia pepper (Capsicum chinense Jacq.) plantations in Assam, India. To investigate possible means to induce plant resistance against this virus, the crude extract of bacterially-expressed double-stranded (ds) RNA, derived from CMV-2b gene (dsRNA_CMV-2b), was exogenously applied along with CMV-G strain onto Bhut Jolokia plants. In this ‘RNA-vaccination’ bioassay, disease incidence, assessed by testing the plants at 21 days post inoculation by DAS-ELISA, ranged from 0 to 29% in case of dsRNA-treated plants, and from 55 to 92% when only CMV was applied. CMV-infected pepper plants became severely stunted, having dull light green foliage with leathery appearance, whereas plants receiving dsRNA_CMV-2b exhibited milder symptoms or remained healthy. The results obtained suggest that this non-transgenic approach has a considerable effect in protecting pepper against CMV.

Keywords

CMV CMV-2b Doubled stranded RNA ELISA RNA vaccine RNAi 

Notes

Acknowledgements

Munmi Borah was supported by a fellowship from European Commission (Erasmus Mundus BRAVE, Grant 2013-2536). Mr Anastasios Katsileros is acknowledged for his assistance in the statistical analysis of the data with JMP. The dsRNA molecules were produced in the frame of a bilateral project between Greece and China (‘sRNAvac’, Grant 12CHN116) funded by the General Secretariat for Research and Technology (GSRT) in Greece.

Supplementary material

13337_2018_452_MOESM1_ESM.docx (186 kb)
Supplementary material 1 (DOCX 185 kb)
13337_2018_452_MOESM2_ESM.docx (1.2 mb)
Supplementary material 2 (DOCX 1232 kb)
13337_2018_452_MOESM3_ESM.docx (16 kb)
Supplementary material 3 (DOCX 15 kb)

References

  1. 1.
    Badu B, Dankers H, Paret ML. First report of Cucumber mosaic virus associated with Capsicum chinense var. Scotch Bonnet in Florida. Plant Dis. 2014;98:1016.Google Scholar
  2. 2.
    Baruah BR, Kashyap A, Nath PD. Incidence, detection and integrated management of viral disease complex in Bhut Jolokia, a chilli cultivar in Assam. Ann Plant Prot Sci. 2016;24:136–41.Google Scholar
  3. 3.
    Bosland PW, Baral JP. Bhut Jolokia—the world’s hottest known chilli is a putative naturally occurring inter specific hybrid. HortScience. 2007;42:222–4.Google Scholar
  4. 4.
    Canto T, Prior DAM, Hellwald K-H, Oparka KJ, Palukaitis P. Characterization of Cucumber Mosaic Virus. IV. Movement protein and coat protein are both essential for cell-to-cell movement of Cucumber mosaic virus. Virology. 1997;237:237–48.CrossRefPubMedGoogle Scholar
  5. 5.
    Dalmay T, Hamilton A, Rudd S, Angell S, Baulcombe DC. An RNA-dependent RNA polymerase gene in Arabidopsis is required for posttranscriptional gene silencing mediated by a transgene but not by a virus. Cell. 2000;101:543–53.CrossRefPubMedGoogle Scholar
  6. 6.
    Ding SW, Li WX, Symons RH. A novel naturally occurring hybrid gene encoded by a plant RNA virus facilitates long distance virus movement. EMBO J. 1995;14:5762–72.PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    González I, Martínez L, Rakitina DV, Lewsey MG, Atencio FA, Llave C, Kalinina NO, Carr JP, Palukaitis P, Canto T. Cucumber mosaic virus 2b protein subcellular targets and interactions: their significance to RNA silencing suppressor activity. Mol Plant Microbe Interact. 2010;23:294–303.CrossRefPubMedGoogle Scholar
  8. 8.
    Holeva MC, Sclavounos AP, Milla SP, Kyriakopoulou PE, Voloudakis AE. External application of dsRNA of the capsid protein (CP) or 2b gene of CMV reduces the severity of CMV-infection in tobacco. In: ΧΙΙΙ International Congress on molecular plant–microbe interactions, 21–27 July 2007, Sorrento, Italy.Google Scholar
  9. 9.
    Meister G, Tuschl T. Mechanisms of gene silencing by double-stranded RNA. Nature. 2004;431:343–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Montgomery MK, Xu S, Fire A. RNA as a target of double-stranded RNA-mediated genetic interference in Caenorhabditis elegans. Proc Natl Acad Sci USA. 1998;95:15502–7.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Naresh P, Reddy MK, Reddy PHC, Reddy KM. Screening chilli (Capsicum spp.) germplasm against Cucumber mosaic virus and Chilli veinal mottle virus and inheritance of resistance. Eur J Plant Pathol. 2016;146:451–64.CrossRefGoogle Scholar
  12. 12.
    Sclavounos AP, Voloudakis AE, Arabatzis C, Kyriakopoulou PE. A severe hellenic CMV tomato isolate: symptom variability in tobacco, characterization and discrimination of variants. Eur J Plant Pathol. 2006;115:63–172.CrossRefGoogle Scholar
  13. 13.
    Sijen T, Kooter JM. Post-transcriptional gene-silencing: RNAs on the attack or on the defense? BioEssays. 2000;22:520–31.CrossRefPubMedGoogle Scholar
  14. 14.
    Suzuki K, Kuroda T, Miura Y, Murai J. Screening and field trials of virus resistant sources in Capsicum spp. Plant Dis. 2003;87:779–83.CrossRefGoogle Scholar
  15. 15.
    Talukdar J, Saikia AK, Borah P. Survey and detection of the diseases of Bhut Jolokia (Capsicum chinense Jacq.) in Assam. J Crop Weed. 2015;11:186–92.Google Scholar
  16. 16.
    Vance V, Vaucheret H. RNA silencing in plants-defense and counterdefense. Science. 2001;292:2277–80.CrossRefPubMedGoogle Scholar
  17. 17.
    Voloudakis AE, Holeva MC, Sarin LP, Bamford DH, Vargas M, Poranen MM, Tenllado F. Efficient double-stranded RNA production methods for utilization in plant virus control. Mol Plant Microbe Interact. 2015;1236:255–74.Google Scholar
  18. 18.
    Zhang X, Yuan YR, Pei Y, Lin SS, Tuschl T, Patel DJ, Chua NH. Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense. Genes Dev. 2006;20:3255–68.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Zitter TA, Murphy JF. Cucumber mosaic. Plant Health Instr. 2009.  https://doi.org/10.1094/PHI-I-2009-0518-01.CrossRefGoogle Scholar

Copyright information

© Indian Virological Society 2018

Authors and Affiliations

  1. 1.Department of Plant PathologyAssam Agricultural UniversityJorhatIndia
  2. 2.Laboratory of Plant Breeding and Biometry, Department of Crop ScienceAgricultural University of AthensAthensGreece
  3. 3.Laboratory of BacteriologyBenaki Phytopathological InstituteKifissiaGreece

Personalised recommendations