Diversity in Betasatellites Associated with Cotton Leaf Curl Disease During Source-To-Sink Movement Through a Resistant Host

Khan, Iftikhar Ali;Akhtar, Khalid Pervaiz;Akbar, Fazal;Hassan, Ishtiaq;Amin, Imran;Saeed, Muhammad;Mansoor, Shahid

  • Received : 2015.08.14
  • Accepted : 2015.11.04
  • Published : 2016.02.01


Cotton leaf curl is devastating disease of cotton characterized by leaf curling, vein darkening and enations. The disease symptoms are induced by DNA satellite known as Cotton leaf curl Multan betasatellite (CLCuMuB), dominant betasatellite in cotton but another betasatellite known as Chili leaf curl betasatellite (ChLCB) is also found associated with the disease. Grafting experiment was performed to determine if host plant resistance is determinant of dominant population of betasatellite in cotton (several distinct strains of CLCuMuB are associated with the disease). Infected scion of Gossypium hirsutum collected from field (the source) was grafted on G. arboreum, a diploid cotton species, resistant to the disease. A healthy scion of G. hirsutum (sink) was grafted at the top of G. arboreum to determine the movement of virus/betasatellite to upper susceptible scion of G. hirsutum. Symptoms of disease appeared in the upper scion and presence of virus/betasatellite in the upper scion was confirmed via molecular techniques, showing that virus/betasatellite was able to move to upper scion through resistant G. arboreum. However, no symptoms appeared on G. arboreum. Betasatelites were cloned and sequenced from lower scion, upper scion and G. arboreum which show that the lower scion contained both CLCuMuB and ChLCB, however only ChLCB was found in G. arboreum. The upper scion contained CLCuMuB with a deletion of 78 nucleotides (nt) in the non-coding region between Arich sequence and ${\beta}C1$ gene and insertion of 27 nt in the middle of ${\beta}C1$ ORF. This study may help in investigating molecular basis of resistance in G. arboreum.


betasatellite;CLCuMuB;Gossypium arboreum;Gossypium hirsutum


  1. Akhtar, K. P., Haidar, S., Khan, M. K. R., Ahmad, M., Sarwar, N., Murtaza, M. A. and Aslam, M. 2010. Evaluation of Gossypium species for resistance to cotton leaf curl burewala virus. Ann. App. Biol. 157:135-147.
  2. Akhtar, K., Ullah, R., Khan, I. A., Saeed, M., Sarwar, N. and Mansoor, S. 2013. First symptomatic evidence of infection of Gossypium arboreum with cotton leaf curl burewala virus through grafting. Int. J. Agri. Bio. 15:157-160.
  3. Akhtar, K. P., Hussain, M., Hassan, M., Sarwar, M. and Sarwar, N. 2015. Evaluation of Bt-cotton genotypes for resistance to cotton leaf curl disease under high inoculum pressure in the field and using graft inoculation in glasshouse. Plant Pathol. J. 3: 132-139.
  4. Amin, I., Mansoor, S., Amrao, L., Hussain, M., Irum, S., Zafar, Y., Bull, S. E. and Briddon, R. W. 2006. Mobilisation into cotton and spread of a recombinant cotton leaf curl disease satellite. Arch. Virol. 151:2055-2065.
  5. Amrao, L., Amin, I., Shahid, M. S., Briddon, R.W. and Mansoor, S. 2010. Cotton leaf curl disease in resistant cotton is associated with a single begomovirus that lacks an intact transcriptional activator protein. Virus. Res. 152:153-163.
  6. Bian, X. Y., Rasheed, M. S., Seemanpillai, M. J. and Rezaian, M. A. 2006. Analysis of silencing escape of Tomato leaf curl virus: an evaluation of the role of DNA methylation. Mol. Plant-Microbe Interact. 19:614-624.
  7. Briddon, R. W. and Stanley, J. 2006. Sub-viral agents associated with plant-infecting single-stranded DNA viruses. Virol. 344:198-210.
  8. Briddon, R. W., Bull, S. E., Amin, I., Mansoor, S., Bedford, I. D., Rishi, N., Siwatch, S. S., Zafar, Y., Abdel-Salam, A. M. and Markham, P. G. 2004. Diversity of DNA 1: a satellite-like molecule associated with monopartite begomovirus-DNA ${\beta}$ complexes. Virology 324:462–74.
  9. Briddon, R. W., Bull, S., Mansoor, S., Amin, I. and Markham, P. G. 2002. Universal primers for the PCR-mediated amplification of DNA ${\beta}$; a satellite associated with some monopartite begomoviruses. Mol. Biotech. 20:315-318.
  10. Doyle, J. J. and Doyle, J. L. 1990. Isolation of plant DNA from fresh tissue. Focus 12:13-15.
  11. Haible, D., Kober, S. and Jeske, H. 2006. Rolling circle amplification revolutionize diagnosis and genomics of geminiviruses. J. Virol. Meth. 135:9-16
  12. Iqbal, Z., Sattar, M. N., Kvarnheden, A., Mansoor, S. and Briddon, R. W. 2012. Effects of the mutation of selected genes of Cotton leaf curl Kokhran virus on infectivity, symptoms and the maintenance of Cotton leaf curl Multan betasatellite. Virus. Res. 169:107-116.
  13. Jeske, H. 2009. Geminiviruses. Cur. Top. Micro. 331:185-226.
  14. Jin, M., Li, C., Shi, Y., Ryabov, E., Huang, J., Wu, Z., Fan, Z. and Hong, Y. 2008. A single amino acid change in a geminiviral Rep protein differentiates between triggering a plant defence response and initiating viral DNA replication. J. Gen. Virol. 89:2636-2641.
  15. Liu, H., Boulton, M. I., Thomas, C. L., Prior, D. A. M., Oparka, K. J. and Davies, J. W. 1999. Maize streak virus coat protein is karyophyllic and facilitates nuclear transport of viral DNA. Mol. Plant-Microbe Interact. 12:894-900.
  16. Mansoor, S., Amin, I., Hussain, M., Zafar, Y. and Briddon, R. W. 2006. Engineering novel traits in plants through RNA interference. Tren. Plant. Sci. 11:559-565.
  17. Mansoor, S., Briddon, R. W., Bull, S. E., Bedford, I. D., Bashir, A., Hussain, M., Saeed, M., Zafar, Y., Malik, K. A., Fauquet, C. and Markham, P. G. 2003. Cotton leaf curl disease is associated with multiple monopartite begomoviruses supported by single DNA. Arch. Virol. 148:1969-1986.
  18. Noueiry, A. O., Lucas, W. J. and Gilbertson, R. L. 1994. Two proteins of a plant DNA virus coordinate nuclear and plasmodesmal transport. Cell 76:925-932.
  19. Saeed, M., Zafar, Y., Randles, J. W. and Rezaian, M. A. 2007. A monopartite begomovirus-associated DNA ${\beta}$ satellite substitutes for the DNA B of a bipartite begomovirus to permit systemic infection. J. Gen. Virol. 88:2881-2889.
  20. Siddique, Z., Akhtar, K. P., Hameed, A., Sarwar, N., Haq, I. U. and Khan, S. A. 2014. Biochemical alterations in leaves of resistant and susceptible cotton genotypes infected systemically by Cotton leaf curl Burewala virus. J. Plant Inter. 9:702-711.
  21. Siddique, Z., Akhtar, K. P., Hameed, A., Haq, I., Ashraf, M. Y., Sarwar, N. and Khan, M. K. R. 2015. Physiological response of cotton leaf curl Burewala virus-infected plants of tolerant and susceptible genotypes of different Gossypium species. J. Plant Pathol. 97:483-490.
  22. Ullah, R., Akhtar, K. P., Moffett, P., Mansoor, S., Briddon, R. W. and Saeed, M. 2014. An analysis of the non-host resistance of Gossypium arboreum to cotton leaf curl disease by grafting. Eur. J. Plant Pathol. 139:837-847.
  23. Wege, C. and Pohl, D. 2007. Abutilon mosaic virus DNA B component supports mechanical virus transmission, but does not counteract begomoviral phloem limitation in transgenic plants. Virol. 365:173-186.
  24. Yun, B. W., Atkinson, H. A., Gaborit, C., Greenland, A., Read, N. D., Pallas, J. A. and Loake, G. J. 2003. Loss of actin cytoskeletal function and EDS1 activity, in combination, severely compromises non-host resistance in Arabidopsis against wheat powdery mildew. Plant J. 34:768-777.

Cited by

  1. Association of chili leaf curl betasatellite with tomato leaf curl disease vol.83, pp.6, 2017,