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Virology Journal

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Open Access 01-12-2015 | Research

RNA interference-based resistance in transgenic tomato plants against Tomato yellow leaf curl virus-Oman (TYLCV-OM) and its associated betasatellite

Authors: Um e Ammara, Shahid Mansoor, Muhammad Saeed, Imran Amin, Rob W Briddon, Abdullah Mohammed Al-Sadi

Published in: Virology Journal | Issue 1/2015

Abstract

Background

Tomato yellow leaf curl virus (TYLCV), a monopartite begomovirus (family Geminiviridae) is responsible for heavy yield losses for tomato production around the globe. In Oman at least five distinct begomoviruses cause disease in tomato, including TYLCV. Unusually, TYLCV infections in Oman are sometimes associated with a betasatellite (Tomato leaf curl betasatellite [ToLCB]; a symptom modulating satellite). RNA interference (RNAi) can be used to develop resistance against begomoviruses at either the transcriptional or post-transcriptional levels.

Results

A hairpin RNAi (hpRNAi) construct to express double-stranded RNA homologous to sequences of the intergenic region, coat protein gene, V2 gene and replication-associated gene of Tomato yellow leaf curl virus-Oman (TYLCV-OM) was produced. Initially, transient expression of the hpRNAi construct at the site of virus inoculation was shown to reduce the number of plants developing symptoms when inoculated with either TYLCV-OM or TYLCV-OM with ToLCB-OM to Nicotiana benthamiana or tomato. Solanum lycopersicum L. cv. Pusa Ruby was transformed with the hpRNAi construct and nine confirmed transgenic lines were obtained and challenged with TYLCV-OM and ToLCB-OM by Agrobacterium-mediated inoculation. For all but one line, for which all plants remained symptomless, inoculation with TYLCV-OM led to a proportion (≤25%) of tomato plants developing symptoms of infection. For inoculation with TYLCV-OM and ToLCB-OM all lines showed a proportion of plants (≤45%) symptomatic. However, for all infected transgenic plants the symptoms were milder and virus titre in plants was lower than in infected non-transgenic tomato plants.

Conclusions

These results show that RNAi can be used to develop resistance against geminiviruses in tomato. The resistance in this case is not immunity but does reduce the severity of infections and virus titer. Also, the betasatellite may compromise resistance, increasing the proportion of plants which ultimately show symptoms.

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Anonymous. Statistics Division of the Food and Agricultural Organization of the United Nations. Available at: http://faostat.fao.org/. Accessed March 2014.

Briddon RW, Brown JK, Moriones E, Stanley J, Zerbini M, Zhou X, et al. Recommendations for the classification and nomenclature of the DNA-β satellites of begomoviruses. Arch Virol. 2008;153:763–81.CrossRefPubMed

Cohen S, Harpaz I. Periodic, rather than continual acquisition of a tomato virus by its vector, the tobacco whitefly (Bemisiatabaci Gennadius). Entomol Exp Appl. 1964;7:155–66.CrossRef

Lefeuvre P, Martin DP, Harkins G, Lemey P, Gray AJ, Meredith S, et al. The spread of tomato yellow leaf curl virus from the Middle East to the world. PLoS Pathog. 2010;6:e1001164.CrossRefPubMedCentralPubMed

Rojas MR, Jiang H, Salati R, Xoconostle-Cazares B, Sudarshana MR, Lucas WJ, et al. Functional analysis of proteins involved in movement of the monopartite begomovirus, Tomato yellow leaf curl virus. Virology. 2001;291:110–25.CrossRefPubMed

Sharma P, Ikegami M. Tomato leaf curl Java virus V2 protein is a determinant of virulence, hypersensitive response and suppression of posttranscriptional gene silencing. Virology. 2010;396:85–93.CrossRefPubMed

Amin I, Hussain K, Akbergenov R, Yadav JS, Qazi J, Mansoor S, et al. Suppressors of RNA Silencing encoded by the components of the cotton leaf curl begomovirus-betasatellite complex. Mol Plant-Microbe Interact. 2011;24:973–83.CrossRefPubMed

Zhang J, Dong J, Xu Y, Wu J. V2 protein encoded by Tomato yellow leaf curl China virus is an RNA silencing suppressor. Virus Res. 2012;163:51–8.CrossRefPubMed

Wang B, Li F, Huang C, Yang X, Qian Y, Xie Y, et al. V2 of tomato yellow leaf curl virus can suppress methylation-mediated transcriptional gene silencing in plants. J Gen Virol. 2014;95:225–30.CrossRefPubMed

10.

Gopal P, Pravin Kumar P, Sinilal B, Jose J, Kasin Yadunandam A, Usha R. 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. 2007;123:9–18.CrossRefPubMed

11.

Luna AP, Morilla G, Voinnet O, Bejarano ER. Functional analysis of gene-silencing suppressors from tomato yellow leaf curl disease viruses. Mol Plant-Microbe Interact. 2012;25:1294–306.CrossRefPubMed

12.

Saeed M, Mansoor S, Rezaian MA, Briddon RW, Randles JW. Satellite DNA beta overrides the pathogenicity phenotype of the C4 gene of tomato leaf curl virus but does not compensate for loss of function of the coat protein and V2 genes. Arch Virol. 2008;153:1367–72.CrossRefPubMed

13.

Hanley-Bowdoin L, Settlage SB, Orozco BM, Nagar S, Robertson D. Geminviruses: models for plant DNA replication, transcription, and cell cycle regulation. Crit Rev Plant Sci. 1999;18:71–106.CrossRef

14.

Laufs J, Traut W, Heyraud F, Matzeit V, Rogers SG, Schell J, et al. In vitro cleavage and joining at the viral origin of replication by the replication initiator protein of tomato yellow leaf curl virus. Proc Natl Acad Sci U S A. 1995;92:3879–83.CrossRefPubMedCentralPubMed

15.

Briddon RW, Mansoor S. Beta ssDNA satellites. In: Mahy BWJ, Van Regenmortel MHV, editors. Encyclopedia of Virology. 3rd ed. Oxford: Elsevier; 2008. p. 314–21.CrossRef

16.

Saunders K, Briddon RW, Stanley J. 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. 2008;89:3165–72.CrossRefPubMed

17.

Cui X, Tao X, Xie Y, Fauquet CM, Zhou X. A DNA β associated with Tomato yellow leaf curl China virus is required for symptom induction. J Virol. 2004;78:13966–74.CrossRefPubMedCentralPubMed

18.

Khan AJ, Idris AM, Al-Saady NA, Al-Mahruki MS, Al-Subhi AM, Brown JK. A divergent isolate of tomato yellow leaf curl virus from Oman with an associated DNA beta satellite: an evolutionary link between Asian and the Middle Eastern virus-satellite complexes. Virus Genes. 2008;36:169–76.CrossRefPubMed

19.

Lecoq H, Moury B, Desbiez C, Palloix A, Pitrat M. Durable virus resistance in plants through conventional approaches: a challenge. Virus Res. 2004;100:31–9.CrossRefPubMed

20.

Foolad MR: Genome mapping and molecular breeding in tomato. Int J Plant Genomics. 2007. article ID 64358. doi:10.1155/2007/64358.

21.

Prins M, Laimer M, Noris E, Schubert J, Wassenegger M, Tepfer M. Strategies for antiviral resistance in transgenic plants. Mol Plant Pathol. 2008;9:73–83.PubMed

22.

Reddy RV, Achenjangb F, Feltona C, Etarocka MT, Anangfaca M, Nugenta P, et al. Role of a geminivirus AV2 protein putative protein kinase C motif on subcellular localization and pathogenicity. Virus Res. 2008;135:115–24.CrossRef

23.

Ding S-W, Voinnet O. Antiviral immunity directed by small RNAs. Cell. 2007;130:413–26.CrossRefPubMedCentralPubMed

24.

Ruiz-Ferrer V, Voinnet O. Roles of plant small RNAs in biotic stress responses. Ann Rev Plant Biol. 2009;60:485–510.CrossRef

25.

Hannon GJ. RNA interference. Nature. 2002;418:244–51.CrossRefPubMed

26.

Baulcombe D. RNA silencing. Trends Biochem Sci. 2005;30:290.CrossRefPubMed

27.

Helliwell CA, Waterhouse PM, Engelke DR, Rossi JJ. Constructs and methods for hairpin RNA-mediated gene silencing in plants. Method Enzymol. 2005;392:24–35.CrossRef

28.

Baulcombe D. RNA silencing in plants. Nature. 2004;431:356–63.CrossRefPubMed

29.

Voinnet O. RNA silencing as a plant immune system against viruses. Trends Genet. 2001;17:449–59.CrossRefPubMed

30.

Chellappan P, Vanitharani R, Fauquet CM. Short interfering RNA accumulation correlates with host recovery in DNA virus-infected hosts, and gene silencing targets specific viral sequences. J Virol. 2004;78:7465–77.CrossRefPubMedCentralPubMed

31.

Lucioli A, Noris E, Brunetti A, Tavazza R, Ruzza V, Castillo AG, et al. Tomato yellow leaf curl Sardinia virus rep-derived resistance to homologous and heterologous geminiviruses occurs by different mechanisms and is overcome if virus-mediated transgene silencing is activated. J Virol. 2003;77:6785–98.CrossRefPubMedCentralPubMed

32.

Mette MF, Aufsatz W, van der Winden J, Matzke MA, Matzke AJ. Transcriptional silencing and promoter methylation triggered by double-stranded RNA. EMBO J. 2000;19:5194–201.CrossRefPubMedCentralPubMed

33.

Pooggin M, Shivaprasad PV, Veluthambi K, Hohn T. RNAi targeting of DNA virus in plants. Nat Biotechnol. 2003;21:131–2.CrossRefPubMed

34.

Aragao FJL, Faria JC. First transgenic geminivirus-resistant plant in the field. Nat Biotechnol. 2009;27:1086–8.CrossRefPubMed

35.

Aragao FJL, Nogueira EO, Tinoco ML, Faria JC. Molecular characterization of the first commercial transgenic common bean immune to the Bean golden mosaic virus. J Biotechnol. 2013;166:42–50.CrossRefPubMed

36.

Aragão FJL, Ribeiro SG, Barros LMG, Brasileiro AM, Maxwell DP, Rech EL, et al. Transgenic beans (Phaseolus vulgaris L.) engineered to express viral antisense RNAs show delayed and attenuated symptoms to bean golden mosaic geminivirus. Mol Breeding. 1998;4:491–9.CrossRef

37.

Nahid N, Amin I, Briddon RW, Mansoor S. RNA interference based resistance against a legume mastrevirus. Virol J. 2011;8:499.CrossRefPubMedCentralPubMed

38.

Lin C-Y, Tsai W-S, Ku H-M, Jan F-J: Evaluation of DNA fragments covering the entire genome of a monopartite begomovirus for induction of viral resistance in transgenic plants via gene silencing. Transgenic Res 2011:1-11.

39.

Saunders K, Bedford ID, Briddon RW, Markham PG, Wong SM, Stanley J. A unique virus complex causes Ageratum yellow vein disease. Proc Natl Acad Sci U S A. 2000;97:6890–5.CrossRefPubMedCentralPubMed

40.

Briddon RW, Mansoor S, Bedford ID, Pinner MS, Saunders K, Stanley J, et al. Identification of DNA components required for induction of cotton leaf curl disease. Virology. 2001;285:234–43.CrossRefPubMed

41.

Cui X, Li G, Wang D, Hu D, Zhou X. A begomovirus DNA β-encoded protein binds DNA, functions as a suppressor of RNA silencing, and targets the cell nucleus. J Virol. 2005;79:10764–75.CrossRefPubMedCentralPubMed

42.

Saeed M, Zafar Y, Randles JW, Rezaian MA. A monopartite begomovirus-associated DNA β satellite substitutes for the DNA B of a bipartite begomovirus to permit systemic infection. J Gen Virol. 2007;88:2881–9.CrossRefPubMed

43.

Mubin M, Hussain M, Briddon RW, Mansoor S. Selection of target sequences as well as sequence identity determine the outcome of RNAi approach for resistance against cotton leaf curl geminivirus complex. Virol J. 2011;8:122.CrossRefPubMedCentralPubMed

44.

Saunders K, Norman A, Gucciardo S, Stanley J. The DNA β satellite component associated with ageratum yellow vein disease encodes an essential pathogenicity protein (βC1). Virology. 2004;324:37–47.CrossRefPubMed

45.

Bonfim K, Faria JC, Nogueira EO, Mendes EA, Aragao FJ. RNAi-mediated resistance to Bean golden mosaic virus in genetically engineered common bean (Phaseolus vulgaris). Mol Plant-Microbe Interact. 2007;20:717–26.CrossRefPubMed

46.

Pelissier T, Thalmeir S, Kempe D, Sanger H, Wassenegger M. Heavy de novo methylation at symmetrical and non-symmetrical sites is a hallmark of RNA-directed DNA methylation. Nucl Acids Res. 1999;27:1625–34.CrossRefPubMedCentralPubMed

47.

Waterhouse PM, Wang M-B, Lough T. Gene silencing as an adaptive defence against viruses. Nature. 2001;411:834–42.CrossRefPubMed

48.

Li F, Huang C, Li Z, Zhou X. Suppression of RNA silencing by a plant DNA virus satellite requires a host calmodulin-like protein to repress RDR6 expression. PLoS Pathog. 2014;10:e1003921.CrossRefPubMedCentralPubMed

49.

Rodríguez-Negrete E, Lozano-Durán R, Piedra-Aguilera A, Cruzado L, Bejarano ER, Castillo AG. Geminivirus Rep protein interferes with the plant DNA methylation machinery and suppresses transcriptional gene silencing. New Phytol. 2013;199:464–75.CrossRefPubMed

50.

Buchmann RC, Asad S, Wolf JN, Mohannath G, Bisaro DM. Geminivirus AL2 and L2 proteins suppress transcriptional gene silencing and cause genome-wide reductions in cytosine methylation. J Virol. 2009;83:5005–13.CrossRefPubMedCentralPubMed

51.

Prins M, Resende RO, Anker C, van Schepen A, De Haan P, Goldbach R. Engineered RNA-mediated resistance to Tomato spotted wilt virus is sequence specific. Mol Plant-Microbe Interact. 1996;9:416–8.CrossRefPubMed

52.

Khan AJ, Akhtar S, Singh AK, Al-Shehi AA, Al-Matrushi AM, Ammara U, et al. Recent evolution of a novel begomovirus causing tomato leaf curl disease in the Al-Batinah region of Oman. Arch Virol. 2014;159:445–55.CrossRefPubMed

53.

Al-Shihi AAM, Khan AJ, Akhtar S, Lima ATM, Zerbini FM, Briddon RW. Occurrence of new recombinant begomovirus species infecting tomato in the Al Batinah region of Oman. Plant Pathol. 2014;63:1177–84.CrossRef

54.

Khan AJ, Akhtar S, Singh AK, Briddon RW. A distinct strain of Tomato leaf curl Sudan virus causes tomato leaf curl disease in Oman. Plant Dis. 2013;97:1396–402.CrossRef

55.

Khan AJ, Akhtar S, Al-Zaidi AM, Singh AK, Briddon RW. Genetic diversity and distribution of a distinct strain of Chili leaf curl virus and associated betasatellite infecting tomato and pepper in Oman. Virus Res. 2013;177:87–97.CrossRefPubMed

56.

Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM. pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation. Plant Mol Biol. 2000;42:819–32.CrossRefPubMed

57.

Ammara U, Maskari AYA, Khan AJ, Al-Sadi AM. Enhanced somatic embryogenesis and Agrobacterium mediated transformation of three cultivars of tomato by exogenous application of putrescine. Int J Agric Biol. 2014;16(1):81–8.

58.

Doyle JJ, Doyle JL. Isolation of plant DNA from fresh tissue. Focus. 1990;12:13–5.

Title: RNA interference-based resistance in transgenic tomato plants against Tomato yellow leaf curl virus-Oman (TYLCV-OM) and its associated betasatellite
Authors: Um e Ammara
Shahid Mansoor
Muhammad Saeed
Imran Amin
Rob W Briddon
Abdullah Mohammed Al-Sadi
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: Virology Journal / Issue 1/2015
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-015-0263-y

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Background

Results

Conclusions

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