European Journal of Plant Pathology

, Volume 123, Issue 2, pp 241–246 | Cite as

Suppression of Papaya ringspot virus infection in Carica papaya with CAP-34, a systemic antiviral resistance inducing protein from Clerodendrum aculeatum

  • Aparana Srivastava
  • Sonali Trivedi
  • Sunil Kumar Krishna
  • H. N. Verma
  • Vivek Prasad


CAP-34, a protein from Clerodendrum aculeatum inducing systemic antiviral resistance was evaluated for control of Papaya ringspot virus (PRSV) infection in Carica papaya. In control plants (treated with CAP-34 extraction buffer) systemic mosaic became visible around 20 days that intensified up to 30 days in 56% plants. During this period, CAP-34-treated papaya did not show any symptoms. Between 30 and 60 days, 95% control plants exhibited symptoms ranging from mosaic to filiformy. In the treated set during the same period, symptoms appeared in only 10% plants, but were restricted to mild mosaic. Presence of PRSV was determined in induced-resistant papaya at the respective observation times by bioassay, plate ELISA, immunoblot and RT-PCR. Back-inoculation with sap from inoculated resistant plants onto Chenopodium quinoa did not show presence of virus. The difference between control and treated sets was also evident in plate-ELISA and immunoblot using antiserum raised against PRSV. PRSV RNA was not detectable in treated plants that did not show symptoms by RT-PCR. Control plants at the same time showed a high intensity band similar to the positive control. We therefore suggest that the absence/delayed appearance of symptoms in treated plants could be due to suppressed virus replication.


Induced systemic resistance Inhibition of virus replication RT-PCR Immunoblot ELISA 



The authors gratefully acknowledge Dr R K Jain, Indian Agricultural Research Institute for the gift of the PRSV antiserum. Financial assistance from the Department of Science and Technology, New Delhi, is also gratefully acknowledged.


  1. Chen, G., Ye, C. M., Huang, J. C., Yu, M., & Li, B. J. (2001). Cloning of the Papaya ringspot virus (PRSV) replicase gene and generation of PRSV-resistant papayas through the introduction of the PRSV replicase gene. Plant Cell Reports, 20, 272–277. doi: 10.1007/s002990000283.CrossRefGoogle Scholar
  2. Chiang, C. H., Lee, C. Y., Wang, C. H., Jan, F. J., Lin, S. S., Chen, T. C., et al. (2007). Genetic analysis of an attenuated Papaya ringspot virus strain applied for cross-protection. European Journal of Plant Pathology, 118, 333–348. doi: 10.1007/s10658-007-9130-z.CrossRefGoogle Scholar
  3. Clark, M. F., & Adam, A. N. (1977). Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant virus. The Journal of General Virology, 34, 475–483. doi: 10.1099/0022-1317-34-3-475.PubMedCrossRefGoogle Scholar
  4. Gonsalves, D. (1998). Control of Papaya ringspot virus in papaya: A case study. Annual Review of Phytopathology, 36, 415–437. doi: 10.1146/annurev.phyto.36.1.415.PubMedCrossRefGoogle Scholar
  5. Jain, R. K., Sharma, J., Shivakumar, A. S., Sharma, P. K., Byadgi, A. S., Verma, A. K., et al. (2004). Variability in the coat protein gene of Papaya ringspot virus isolated from multiple locations in India. Archives of Virology, 149, 2435–2442. doi: 10.1007/s00705-004-0392-x.PubMedCrossRefGoogle Scholar
  6. Jensen, D. D. (1949). Papaya virus diseases with special references to papaya ringspot. Phytopathology, 39, 191–211.Google Scholar
  7. Krubphachaya, P., Juricek, M., & Kertbundit, S. (2007). Induction of RNA-mediated resistance to Papaya ringspot virus type W. Journal of Biochemistry and Molecular Biology, 40, 404–411.PubMedGoogle Scholar
  8. Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685. doi: 10.1038/227680a0.PubMedCrossRefGoogle Scholar
  9. Picard, D., Kao, C. C., & Hudak, K. A. (2005). Pokeweed antiviral protein inhibits Brome mosaic virus replication in plant cells. The Journal of Biological Chemistry, 280, 20069–20075. doi: 10.1074/jbc.M413452200.PubMedCrossRefGoogle Scholar
  10. Prasad, V., Srivastava, S., Varsha, , & Verma, H. N. (1995). Two basic proteins isolated from Clerodendrum inerme Gaertn. are inducers of systemic antiviral resistance in susceptible plants. Plant Science, 110, 73–82. doi: 10.1016/0168-9452(95)04192-W.CrossRefGoogle Scholar
  11. Prasad, V., & Srivastava, S. (2001). Inducible mechanisms of plant resistance to virus infection. Journal of Plant Biology, 28, 1–11.Google Scholar
  12. Ritzenthaler, C. (2005). Resistance to plant viruses: old issues, new answers. Current Opinion in Biotechnology, 16, 118–122. doi: 10.1016/j.copbio.2005.02.009.PubMedCrossRefGoogle Scholar
  13. Tennant, P., Ahmad, M. H., & Gonsalves, D. (2005). Field resistance of coat protein transgenic papaya to Papaya ringspot virus in Jamaica. Plant Disease, 89, 841–847. doi: 10.1094/PD-89-0841.CrossRefGoogle Scholar
  14. Towbin, H., Stahelin, T., & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America, 76, 4350–4354. doi: 10.1073/pnas.76.9.4350.PubMedCrossRefGoogle Scholar
  15. Tripathi, S., Bau, H. J., Chen, L. F., & Yeh, S. D. (2004). The ability of Papaya ringspot virus strains overcoming the transgenic resistance of papaya conferred by the coat protein gene is not correlated with higher degrees of sequence divergence from the transgene. European Journal of Plant Pathology, 110, 871–882. doi: 10.1007/s10658-004-0607-8.CrossRefGoogle Scholar
  16. Van Loon, L. C., Bakker, P. A. H. M., & Pieterse, C. M. J. (1998). Systemic resistance induced by rhizophere bacteria. Annual Review of Phytopathology, 36, 453–483. doi: 10.1146/annurev.phyto.36.1.453.PubMedCrossRefGoogle Scholar
  17. Verma, H. N., & Awasthi, L. P. (1979). Antiviral activity of Boerhaavia diffusa root extract and the physical properties of the virus inhibitor. Canadian Journal of Botany, 57, 926–932. doi: 10.1139/b79-113.CrossRefGoogle Scholar
  18. Verma, H. N., & Prasad, V. (1986). Virus diseases in Papaw (papaya). In S. P. Raychaudhuri, & J. P. Verma (Eds.), Review of tropical plant pathology (vol. II, (pp. 311–327)). New Delhi, India: Today and Tomorrows.Google Scholar
  19. Verma, H. N., Srivastava, S., Varsha, , & Kumar, D. (1996). Induction of systemic resistance in plants against viruses by a basic protein from Clerodendrum aculeatum leaves. Phytopathology, 86, 485–492. doi: 10.1094/Phyto-86-485.CrossRefGoogle Scholar
  20. Yang, X., Liangyi, K., & Tien, P. (1996). Resistance of tomato infected with Cucumber mosaic virus satellite RNA to Potato spindle tuber viroid. The Annals of Applied Biology, 129, 543–551. doi: 10.1111/j.1744-7348.1996.tb05775.x.CrossRefGoogle Scholar

Copyright information

© KNPV 2008

Authors and Affiliations

  • Aparana Srivastava
    • 1
  • Sonali Trivedi
    • 1
  • Sunil Kumar Krishna
    • 1
  • H. N. Verma
    • 2
  • Vivek Prasad
    • 1
  1. 1.Molecular Plant Virology Lab, Department of BotanyUniversity of LucknowLucknowIndia
  2. 2.Seedling Academy of Design, Technology and ManagementJaipurIndia

Personalised recommendations