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Applied Microbiology and Biotechnology

, Volume 97, Issue 12, pp 5457–5471 | Cite as

Infection of tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus with betasatellites, results in enhanced level of helper virus components and antagonistic interaction between DNA B and betasatellites

  • P. Jyothsna
  • Q. M. I. Haq
  • Priyanka Singh
  • K. V. Sumiya
  • Shelly Praveen
  • Ramaveer Rawat
  • Rob W. Briddon
  • V. G. Malathi
Applied genetics and molecular biotechnology

Abstract

Tomato leaf curl New Delhi virus (ToLCNDV) (Geminiviridae) is an important pathogen that severely affects tomato production. An extensive survey was carried out during 2003–2010 to study the diversity of begomoviruses found in tomato, potato, and cucurbits that showed symptoms of leaf puckering, distortion, curling, vein clearing, and yellow mosaic in various fields in different regions of India. Ten begomovirus isolates were cloned from infected samples and identified as belonging to the species ToLCNDV. A total of 44 % of the samples showed association of betasatellites, with CLCuMuB and LuLDB being the most frequent. The ToLCNDV cloned component DNA A and DNA B were agroinoculated on Nicotiana benthamiana and tomato (Solanum lycopersicum) plants with or without betasatellites, CLCuMuB or LuLDB. The viral genome levels were then monitored by real-time polymerase chain reaction at different time points of disease development. Plants co-inoculated with betasatellites showed enhanced symptom severity in both N. benthamiana and tomato, as well as increases in helper viral DNA A and DNA B levels. The DNA B and betasatellites acted antagonistically to each other, so that the level of DNA B was 16-fold greater in the presence of betasatellites, while accumulation of betasatellites, CLCuMuB and LuLDB, were reduced by 60 % in the presence of DNA B. DNA B-mediated symptoms predominated in CLCuMuB-inoculated plants, whereas betasatellite-mediated leaf abnormalities were prominent in LuLDB-co-inoculated plants. Inoculation with the cloned components will be a good biotechnological tool in resistance breeding program.

Keywords

Begomovirus Tomato leaf curl New Delhi virus Antagonism DNA B Betasatellites 

Notes

Acknowledgments

The authors are grateful for the facilities provided by Dr. H.S. Gupta, The Director, Indian Agricultural Research Institute and Dr. R.K. Jain, Head, Division of Plant Pathology in conducting this study. The authors are also thankful to Darren P. Martin and Brejnev Muhire, Institute of Infectious Disease and Molecular Medicine, Computational Biology Group, University of Cape Town, South Africa for their guidance in the SDT analysis. The financial assistance by the Department of Biotechnology, Government of India, Project BT/PR/5351/AGR/16/482/2004-II is duly acknowledged. The support of the National Phytotron Facility is highly appreciated.

Supplementary material

253_2012_4685_MOESM1_ESM.pdf (379 kb)
ESM 1 (PDF 378 kb)

References

  1. Amin I, Hussain K, Akbergenov R, Yadav JS, Qazi J, Mansoor S, Hohn T, Fauquet CM, Briddon RW (2011) Suppressors of RNA silencing encoded by the components of the cotton leaf curl begomovirus–betasatellite complex. Mol Plant Microbe Interact 24:973–983CrossRefGoogle Scholar
  2. Bevan MW (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721CrossRefGoogle Scholar
  3. Briddon RW, Stanley J (2006) Subviral agents associated with plant single-stranded DNA viruses. Virology 344:198–210CrossRefGoogle Scholar
  4. Briddon RW, Mansoor S, Bedford ID, Pinner MS, Saunders K, Stanley J, Zafar Y, Malik K, Markham PG (2001) Identification of DNA components required for induction of cotton leaf curl disease. Virology 285:234–243CrossRefGoogle Scholar
  5. Briddon RW, Bull SE, Mansoor S, Amin I, Markham PG (2002) Universal primers for the PCR-mediated amplification of DNA β: a molecule associated with some monopartite begomoviruses. Mol Biol Technol 20:315–318Google Scholar
  6. Briddon RW, Brown JK, Moriones E, Stanley J, Zerbini M, Zhou X, Fauquet CM (2008) Recommendations for the classification and nomenclature of the DNA-β satellites of begomoviruses. Arch Virol 153:763–781CrossRefGoogle Scholar
  7. Bull SE, Tsai WS, Briddon RW, Markham PG, Stanley J, Green SK (2004) Diversity of begomovirus DNA β satellites of non-malvaceous plants in east and south east Asia. Arch Virol 149:1193–1200CrossRefGoogle Scholar
  8. Cui X, Tao X, Xie Y, Fauquet CM, Zhou X (2004) A DNAβ associated with tomato yellow leaf curl China virus is required for symptom induction. J Virol 78:13966–13974CrossRefGoogle Scholar
  9. Cui X, Li G, Wang D, Hu D, Zhou XP (2005) A begomovirus DNA β-encoded protein binds DNA, functions as a suppressor of RNA silencing, and targets the cell nucleus. J Virol 79:10764–10775CrossRefGoogle Scholar
  10. Duan YP, Powell CA, Purcifull DE, Broglio P, Hiebert E (1997) Phenotypic variation in transgenic tobacco expressing mutated geminivirus movement/pathogenicity (BC1) proteins. Mol Plant Microbe Interact 10:1065–1074CrossRefGoogle Scholar
  11. Edgar RC (2004) Muscle: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797CrossRefGoogle Scholar
  12. Fauquet CM, Briddon RW, Brown JK, Moriones E, Stanley J, Zerbini M, Zhou X (2008) Geminivirus strain demarcation and nomenclature. Arch Virol 153:783–821CrossRefGoogle Scholar
  13. Gopal P, Kumar PP, Sinilal B, Jose J, Yadunandam KA, Usha R (2007) Differential roles of C4 and βC1 in mediating suppression of post-transcriptional gene silencing: evidence for transactivation by the C2 of Bhendi yellow vein mosaic virus, a monopartite begomovirus. Virus Res 123:9–18CrossRefGoogle Scholar
  14. Guo W, Jiang T, Zhang X, Li G, Zhou X (2008) Molecular variation of satellite DNA β molecules associated with Malvastrum yellow vein virus and their role in pathogenicity. Appl Environ Microbiol 74:1909–1913CrossRefGoogle Scholar
  15. Guo W, Yang X, Xie Y, Cui X, Zhou X (2009) Tomato yellow leaf curl Thailand virus-[Y72] from Yunnan is a monopartite begomovirus associated with DNAβ. Virus Genes 38:328–333CrossRefGoogle Scholar
  16. Haible D, Kober S, Jeske H (2006) Rolling circle amplification revolutionizes diagnosis and genomics of geminiviruses. J Virol Meth 135:9–16CrossRefGoogle Scholar
  17. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp 41:95–98Google Scholar
  18. Hanley-Bowdoin L, Settlage SB, Orozco BM, Nagar S, Robertson D (1999) Geminiviruses: models for plant DNA replication, transcription and cell cycle regulation. Crit Rev Plant Sci 18:71–106CrossRefGoogle Scholar
  19. Hood EE, Gelvin SB, Melchers S, Hoekema A (1993) New Agrobacterium helper plasmid for gene transfer to plants (EHA 105). Transgenic Res 2:208–218CrossRefGoogle Scholar
  20. Hou YM, Saunders R, Ursin VM, Gilbertson RL (2000) Transgenic plants expressing geminivirus movement proteins: abnormal phenotypes and delayed infection by tomato mottle virus in transgenic tomatoes expressing the bean dwarf mosaic virus BV1 or BC1 proteins. Mol Plant Microbe Interact 13:297–308CrossRefGoogle Scholar
  21. Hussain M, Mansoor S, Iram S, Fatima AN, Zafar Y (2005) The nuclear shuttle protein of tomato leaf curl New Delhi virus is a pathogenicity determinant. J Virol 79:4434–4439CrossRefGoogle Scholar
  22. Ilyas M, Qazi J, Mansoor S, Briddon RW (2010) Genetic diversity and phylogeography of begomoviruses infecting legumes in Pakistan. J Gen Virol 91:2091–2101CrossRefGoogle Scholar
  23. Ingham DJ, Pascal E, Lazarowitz SG (1995) Both bipartite geminivirus movement proteins define viral host range, but only BL1 determines viral pathogenicity. Virology 207:191–204CrossRefGoogle Scholar
  24. Jose J, Usha R (2003) Bhendi yellow vein mosaic disease in India is caused by association of a DNA β satellite with a begomovirus. Virology 305:310–317CrossRefGoogle Scholar
  25. Kings AMQ, Adams MJ, Carstens EB, Lefkowitz EJ (2011) Virus taxonomy. Ninth report of the International Committee on Taxonomy of Viruses. Elsevier Academic Press, San DiegoGoogle Scholar
  26. Kon T, Sharma P, Ikegami M (2007) Suppressor of RNA silencing encoded by the monopartite tomato leaf curl Java begomovirus. Arch Virol 152:1273–1282CrossRefGoogle Scholar
  27. Kumari P, Singh AK, Sharma VK, Chattopadhyay B, Chakraborty S (2011) A novel recombinant tomato-infecting begomovirus capable of trans-complementing heterologous DNA-B components. Arch Virol 156:769–783CrossRefGoogle Scholar
  28. Li ZH, Xie Y, Zhou XP (2005) Tobacco curly shoot virus DNA-β is not necessary for infection but intensifies symptoms in a host-dependent manner. Phytopathol 95:902–908CrossRefGoogle Scholar
  29. Lin B, Behjatnia SAA, Dry IB, Randles JW, Rezaian MA (2003) High-affinity Rep-binding is not required for the replication of a geminivirus DNA and its satellite. Virology 305:353–363CrossRefGoogle Scholar
  30. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔC T Method. Methods 25:402–408CrossRefGoogle Scholar
  31. Mansoor S, Briddon RW, Bull SE, Bedford ID, Bashir A, Hussain M, Saeed M, Zafar MY, Malik KA, Fauquet CM, Markham PG (2003) Cotton leaf curl disease is associated with multiple monopartite begomoviruses supported by single DNA β. Arch Virol 148:1969–1986CrossRefGoogle Scholar
  32. Nawaz-ul-Rehman MS, Mansoor S, Briddon RW, Fauquet CM (2009) Maintenance of an Old World betasatellite by a new world helper begomovirus and possible rapid adaptation of the betasatellite. J Virol 83:9347–9355CrossRefGoogle Scholar
  33. Padidam M, Beachy RN, Fauquet CM (1995) Tomato leaf curl geminivirus from India has a bipartite genome and coat protein is not essential for infectivity. J Gen Virol 76:25–35CrossRefGoogle Scholar
  34. Pascal E, Goodlove PE, Wu LC, Lazarowitz SG (1993) Transgenic tobacco plants expressing the geminivirus BL1 protein exhibit symptoms of viral disease. Plant Cell 5:795–807Google Scholar
  35. Pratap D, Kashikar AR, Mukherjee SK (2011) Molecular characterization and infectivity of a tomato leaf curl New Delhi virus variant associated with newly emerging yellow mosaic disease of eggplant in India. Virology J 8:305CrossRefGoogle Scholar
  36. Qazi J, Ilyas M, Mansoor S, Briddon RW (2007) Legume yellow mosaic viruses: genetically isolated begomoviruses. Mol Plant Pathol 8:343–348CrossRefGoogle Scholar
  37. Rojas MR, Jiang H, Salati R, Xoconostle-Cázares B, Sudarshana MR, Lucas WJ, Gilbertson RL (2001) Functional analysis of proteins involved in movement of the monopartite begomovirus, tomato yellow leaf curl virus. Virology 5:110–125CrossRefGoogle Scholar
  38. Rojas MR, Hagen C, Lucas WJ, Gilbertson RL (2005) Exploiting chinks in the plant’s armor: evolution and emergence of geminiviruses. Ann Rev Phytopathol 43:361–394CrossRefGoogle Scholar
  39. Rouhibakhsh A, Malathi VG (2005) Severe leaf curl disease of cowpea—a new disease of cowpea in northern India caused by mungbean yellow mosaic India virus and a satellite DNA β. Plant Pathol 54:259CrossRefGoogle Scholar
  40. Rouhibakhsh A, Priya J, Periasamy M, Haq QMI, Malathi VG (2008) An improved DNA isolation method and PCR protocol for efficient detection of multicomponents of begomovirus in legumes. J Virol Meth 147:37–42CrossRefGoogle Scholar
  41. Saeed M, Behjatnia SAA, Mansoor S, Zafar Y, Hasnain S, Rezaian MA (2005) A single complementary-sense transcript of a geminiviral DNA-β satellite is determinant of pathogenicity. Mol Plant Microbe Interact 18:7–14CrossRefGoogle Scholar
  42. Saeed M, Zafar Y, Randles JW, Rezaian MA (2007) A monopartite begomovirus-associated DNA β satellite substitutes for the DNA B of a bipartite begomovirus to permit systemic infection. J Gen Virol 88:2881–2889CrossRefGoogle Scholar
  43. Sahu PP, Rai NK, Chakraborty S, Singh M, Chandrappa PH, Ramesh B, Chattopadhyay D, Prasad M (2010) Tomato cultivar tolerant to tomato leaf curl New Delhi virus infection induces virus-specific short interfering RNA accumulation and defence-associated host gene expression. Mol Plant Pathol 11:531–544CrossRefGoogle Scholar
  44. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning a laboratory manual, vol I, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  45. Saunders K, Salim N, Mali VR, Malathi VG, Briddon RW, Markham PG, Stanley J (2002) Characterisation of Sri Lankan cassava mosaic virus and Indian cassava mosaic virus: evidence for acquisition of a DNA B component by a monopartite begomovirus. Virology 293:63–74CrossRefGoogle Scholar
  46. Saunders K, Norman A, Gucciardo S, Stanley J (2004) The DNA β satellite component associated with Ageratum yellow vein disease encodes an essential pathogenicity protein (βC1). Virology 324:37–47CrossRefGoogle Scholar
  47. Saunders K, Briddon RW, Stanley J (2008) Replication promiscuity of DNA-β satellites associated with monopartite begomoviruses; deletion mutagenesis of the Ageratum yellow vein virus DNA-β satellite localizes sequences involved in replication. J Gen Virol 89:3165–3172CrossRefGoogle Scholar
  48. Sharma P, Ikegami M, Kon T (2010) Identification of the virulence factors and suppressors of posttranscriptional gene silencing encoded by Ageratum yellow vein virus, a monopartite begomovirus. Virus Res 149:19–27CrossRefGoogle Scholar
  49. Shimura H, Pantaleo V (2011) Viral induction and suppression of RNA silencing in plants. Biochim Biophys Acta 1809:601–612CrossRefGoogle Scholar
  50. Sivalingam PN, Malathi VG, Varma A (2010) Molecular diversity of the DNA-β satellites associated with tomato leaf curl disease in India. Arch Virol 155:757–764CrossRefGoogle Scholar
  51. Srivastava KM, Hallan V, Raizada RK, Chandra G, Singh BP, Sane PV (1995) Molecular cloning of Indian tomato leaf curl virus genome following a simple method of concentrating the supercoiled replicative form of viral DNA. J Virol Meth 51:297–304CrossRefGoogle Scholar
  52. Stanley J (1985) The molecular biology of geminiviruses. Adv Virus Res 30:139–177CrossRefGoogle Scholar
  53. Stanley J, Townsend R (1985) Characterization of DNA forms associated with cassava latent virus infection. Nucleic Acids Res 13:2189–2206CrossRefGoogle Scholar
  54. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefGoogle Scholar
  55. Tiwari N, Padmalatha KV, Singh VB, Haq QMI, Malathi VG (2010) Tomato leaf curl Bangalore virus (ToLCBV): infectivity and enhanced pathogenicity with diverse betasatellites. Arch Virol 155:1343–1347CrossRefGoogle Scholar
  56. von Arnim A, Stanley J (1992) Determinants of tomato golden mosaic virus symptom development located on DNA B. Virology 186:286–293CrossRefGoogle Scholar
  57. Yang JY, Iwasaki M, Machida C, Machida Y, Zhou X, Chua NH (2008) βC1, the pathogenicity factor of TYLCCNV, interacts with AS1 to alter leaf development and suppress selective jasmonic acid responses. Genes Dev 15:2564–2577CrossRefGoogle Scholar
  58. Yang X, Wang Y, Guo W, Xie Y, Xie Q, Fan L, Zhou X (2011) Characterization of small interfering RNAs derived from the geminivirus/betasatellite complex using deep sequencing. PLoS One 6:e16928CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • P. Jyothsna
    • 1
  • Q. M. I. Haq
    • 1
  • Priyanka Singh
    • 1
  • K. V. Sumiya
    • 1
  • Shelly Praveen
    • 1
  • Ramaveer Rawat
    • 2
  • Rob W. Briddon
    • 3
  • V. G. Malathi
    • 1
  1. 1.Advanced Centre for Plant Virology, Division of Plant PathologyIndian Agricultural Research InstituteNew DelhiIndia
  2. 2.MM (PG) CollegeModinagarIndia
  3. 3.National Institute for Biotechnology and Genetic EngineeringFaisalabadPakistan

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