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Resistance to a DNA and a RNA virus in transgenic plants by using a single chimeric transgene construct

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Abstract

Tomato leaf curl Taiwan virus (ToLCTWV) and Tomato spotted wilt virus (TSWV) are two major tomato viruses that cause serious economic losses. In this study, a partial C2 gene from ToLCTWV and the middle half of the N gene of TSWV were fused as a chimeric transgene to develop multiple virus resistance in transgenic plants. This construct was introduced into Nicotiana benthamiana and tomato by Agrobacterium-mediated transformation. Several transgenic lines showed no symptom post agro-inoculation with ToLCTWV and displayed high resistance to TSWV. The detection of siRNAs indicated that the resistance was via RNA silencing. This study demonstrated that linkage of gene segments from two viruses with distinct genomic organization, one DNA and the other RNA, can confer multiple virus resistance in transgenic plants via gene silencing.

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Abbreviations

PTGS:

Post-transcriptional gene silencing

siRNA:

Small interfering RNA

ToLCTWV:

Tomato leaf curl Taiwan virus

TSWV:

Tomato spotted wilt virus

References

  • Aragão FJL, Ribeiro SG, Barros LMG, Brasileiro ACM, Maxwell DP, Rech EL, Faria JC (1998) Transgenic beans (Phaseolus vulgaris L.) engineered to express viral antisense RNAs show delayed and attenuated symptoms to bean golden mosaic geminivirus. Mol Breed 4:491–499

    Article  Google Scholar 

  • Asad S, Haris WA, Bashir A, Zafar Y, Malik KA, Malik NN, Lichtenstein CP (2003) Transgenic tobacco expressing geminiviral RNAs is resistant to the serious viral pathogen causing cotton leaf curl disease. Arch Virol 148:2341–2352

    Article  CAS  PubMed  Google Scholar 

  • Bai QR, Zhu JH, Liu XL, Zhu CX, Song YZ, Wen FJ (2005) Production of transgenic tobacco plants resistant to two viruses via RNA-mediated virus resistance. Acta Phytopathol Sin 35:148–154

    Google Scholar 

  • Bejarano ER, Lichtenstein CP (1994) Expression of TGMV antisense RNA in transgenic tobacco inhibits replication of BCTV but not ACMV geminiviruses. Plant Mol Biol 24:241–248

    Article  CAS  PubMed  Google Scholar 

  • Bernstein E, Caudy AA, Hammond SM, Hannon GJ (2001) Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409:363–366

    Article  CAS  PubMed  Google Scholar 

  • Bucher E, Lohuis D, Pieter M, van Poppel JA, Geerts-Dimitriadou C, Goldbach R, Prins M (2006) Multiple virus resistance at a high frequency using a single transgene construct. J Gen Virol 87:3697–3701

    Article  CAS  PubMed  Google Scholar 

  • Chellappan P, Masona MV, Vanitharani R, Taylor NJ, Fauquet CM (2004) Broad spectrum resistance to ssDNA viruses associated with transgene-induced gene silencing in cassava. Plant Mol Biol 56:601–611

    Article  CAS  PubMed  Google Scholar 

  • de Haan P, Wagemakers L, Peters D, Goldbach R (1990) The S RNA segment of tomato spotted wilt virus has an ambisense character. J Gen Virol 71:1001–1007

    Article  PubMed  Google Scholar 

  • de Haan P, Gielen JJL, Prins M, Wijkamp IG, van Schepen A, Peters D, van Grinsven MQJM, Goldbach R (1992) Characterization of RNA-mediated resistance to tomato spotted wilt virus in transgenic tobacco plants. Bio/Technology 10:1133–1137

    Google Scholar 

  • Fulton TM, Chunwongse J, Tanksley SD (1995) Microprep protocol for extraction of DNA from tomato and other herbaceous plants. Plant Mol Biol Rep 13:207–209

    Article  CAS  Google Scholar 

  • Gielen JJL, de Haan P, Kool AJ, Peters D, van Grinsven MQJM, Goldbach RW (1991) Engineered resistance to tomato spotted wilt virus, a negative-strand RNA virus. Bio/Technology 9:1363–1367

    Article  CAS  Google Scholar 

  • Hamilton AJ, Baulcombe DC (1999) A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286:950–952

    Article  CAS  PubMed  Google Scholar 

  • Hammond SM, Bernstein E, Beach D, Hannon GJ (2000) RNA directed nuclease mediates posttranscriptional gene silencing in Drosophila cell extracts. Nature 404:293–296

    Article  CAS  PubMed  Google Scholar 

  • Hao L, Wang H, Sunter G, Bisaro DM (2003) Geminivirus AL2 and L2 proteins interact with and inactivate SNF1 kinase. Plant Cell 15:1034–1048

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann HL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1230

    Article  CAS  Google Scholar 

  • Jan F-J (1998) Roles of nontarget DNA and viral gene length in influencing multi-virus resistance through homology-dependent gene silencing. Ph. D. Dissertation, Department of Plant Pathology, Cornell University, Ithaca

  • Jan F-J, Fagoaga C, Pang S-Z, Gonsalves D (2000a) A minimum length of N gene sequence in transgenic plants is required for RNA-mediated tospovirus resistance. J Gen Virol 81:235–242

    CAS  PubMed  Google Scholar 

  • Jan F-J, Fagoaga C, Pang S-Z, Gonsalves D (2000b) A single chimeric transgene derived from two distinct viruses confers multi-virus resistance in transgenic plants through homology-dependent gene silencing. J Gen Virol 81:2103–2109

    Article  CAS  PubMed  Google Scholar 

  • Kasschau KD, Xie Z, Allen E, Lave C, Chapman EJ, Krizan KA, Carrington JC (2003) P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA function. Dev Cell 4:205–217

    Article  CAS  PubMed  Google Scholar 

  • Mansoor S, Briddon RW, Zafar Y, Stanley J (2003) Geminivirus disease complexes: an emerging threat. Trends Plant Sci 8:128–134

    Article  CAS  PubMed  Google Scholar 

  • Napoli C, Lemieux C, Jorgensen R (1990) Introduction of chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2:279–290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nicola-Negri ED, Brunetti A, Tavazza M, Ilardi V (2005) Hairpin RNA-mediated silencing of Plum pox virus P1 and HC-Pro genes for efficient and predictable resistance to the virus. Transgenic Res 14:989–994

    Article  PubMed  CAS  Google Scholar 

  • Niu Q-W, Lin S-S, Reyes JL, Chen K-C, Wu H-W, Yeh S-D, Chua N-H (2006) Expression of artificial microRNAs in transgenic Arabidopsis thaliana confers virus resistance. Nat Biotechnol 24:1420–1428

    Article  CAS  PubMed  Google Scholar 

  • Pang S-Z, Nagpala P, Wang M, Slightom JL, Gonsalves D (1992) Resistance to heterologous isolates of tomato spotted wilt virus in transgenic tobacco expressing its nucleocapsid protein gene. Phytopathology 82:1223–1229

    Article  CAS  Google Scholar 

  • Pang S-Z, Jan F-J, Gonsalves D (1997) Nontarget DNA sequences reduce the transgene length necessary for RNA-mediated tospovirus resistance in transgenic plants. Proc Natl Acad Sci USA 94:8261–8266

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Praveen S, Mishra AK, Antony G (2006) Viral suppression in transgenic plants expression chimeric transgene from tomato leaf curl virus and cucumber mosaic virus. Plant Cell Tissue Organ Cult 84:47–53

    Article  CAS  Google Scholar 

  • Prins M, Goldbach R (1998) The emerging problem of tospovirus infection and nonconventional methods of control. Trends Microbiol 6:31–35

    Article  CAS  PubMed  Google Scholar 

  • Prins M, Resende RO, Anker C, Van Schepen A, de Haan P, Goldbach R (1996) Engineered RNA-mediated resistance to tomato spotted wilt virus is sequence specific. Mol Plant Microbe Interact 9:416–418

    Article  CAS  PubMed  Google Scholar 

  • Qu J, Ye J, Fang R (2007) Artificial microRNA-mediated virus resistance in plants. J Virol 81:6690–6699

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rojas MR, Gilbertson RT, Russell DR, Maxwell DP (1993) Use of degenerate primers in the polymerase chain reaction to detect whitefly-transmitted geminiviruses. Plant Dis 77:340–347

    Article  CAS  Google Scholar 

  • Shih SL, Green SK, Lee LM, Wang JT, Tsai WS, Ledesma DR, Chen JT (2004) On-farm evaluation of tomato leaf curl disease control measures in Taiwan. Plant Prot Bull 46:417–418

    Google Scholar 

  • Sonoda S, Tsumuki H (2004) Analysis of RNA-mediated virus resistance by NSs and NSm gene sequences from Tomato spotted wilt virus. Plant Sci 16:771–778

    Article  CAS  Google Scholar 

  • Sunter G, Bisaro DM (1992) Transactivation of geminivirus AR1 and BR1 gene expression by the AL2 gene product occurs at the level of transcription. Plant Cell 4:1321–1331

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sunter G, Sunter J, Bisaro DM (2001) Plants expressing Tomato golden mosaic virus AL2 or beet curly top virus L2 transgenes show enhanced susceptibility to infection by DNA and RNA viruses. Virology 285:59–70

    Article  CAS  PubMed  Google Scholar 

  • Tsai W-S (2008) Molecular characterization of Chilli veinal mottle virus from South and Southeast Asia and tomato begomoviruses from the Philippines and Taiwan, and development of transgenic resistance to Tomato leaf curl Taiwan virus. Ph. D. Dissertation, Department of Plant Pathology, National Chung Hsing University, Taichung

  • van Wezel R, Liu H, Tien P, Stanley J, Hong Y (2002) Mutation of three cysteine residues in tomato yellow leaf curl virus-China C2 protein causes dysfunction in pathogenesis and posttranscriptional gene silencing-suppression. Mol Plant Microbe Interact 15:203–208

    Article  Google Scholar 

  • Vanitharani R, Chellappan P, Pita JS, Fauquet C (2004) Differential roles of AC2 and AC4 of cassava geminiviruses in mediating synergism and suppression of posttranscriptional gene silencing. J Virol 78:9487–9498

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vanitharani R, Chellappan P, Fauquet CM (2005) Geminiviruses and RNA silencing. Trends Plant Sci 10:144–151

    Article  CAS  PubMed  Google Scholar 

  • Xie Y, Jiang T, Zhou X (2006) Agroinoculation shows Tobacco leaf curl Yunnan virus is a monopartite begomovirus. Eur J Plant Pathol 115:369–375

    Article  Google Scholar 

  • Xue B, Gonsalves C, Provvidenti R, Slightom JL, Fuchs M, Gonsalves D (1994) Development of transgenic tomato expressing a high level of resistance to cucumber mosaic virus strains of subgroups I and II. Plant Dis 78:1038–1041

    Article  Google Scholar 

  • Yeh SD, Gonsalves D (1984) Purification and immunological analyses of cylindrical-inclusion protein induced by papaya ringspot virus and watermelon mosaic virus 1. Phytopathology 74:1273–1278

    Article  CAS  Google Scholar 

  • Zhang P, Vanderschuren H, Futterer J, Gruissem W (2005) Resistance to cassava mosaic disease in transgenic cassava expressing antisense RNAs targeting virus replication genes. Plant Biotechnol J 3:385–397

    Article  PubMed  CAS  Google Scholar 

  • Zhu C-X, Song Y-Z, Yin G-H, Wen F-J (2009) Induction of RNA-mediated multiple virus resistance to Potato virus Y, Tobacco mosaic virus and Cucumber mosaic virus. J Phytopathol 157:101–107

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful to Dr. Wen-Hsiung Ko, Professor Emeritus, University of Hawaii at Manoa, Dr. Chung-Jan Chang, Professor, University of Georgia and Dr. Michael Thomson at International Rice Research Institute (IRRI) for their critical review of this manuscript. We thank Dr. Shyi-Dong Yeh for providing TSWV antibody. This work was supported by a grant (NSC97-2317-B-005-004) of the “National Science and Technology Program for Agricultural Biotechnology” from the National Science Council of Taiwan.

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Authors and Affiliations

  1. Department of Plant Pathology, National Chung Hsing University, Taichung, 402, Taiwan

    Ching-Yi Lin & Fuh-Jyh Jan

  2. Department of Agronomy, National Chung Hsing University, Taichung, 402, Taiwan

    Hsin-Mei Ku

  3. Asian Vegetable Research and Development Center—The World Vegetable Center, Tainan, 741, Taiwan

    Wen-Shi Tsai & Sylvia K. Green

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  1. Ching-Yi Lin
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  2. Hsin-Mei Ku
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  5. Fuh-Jyh Jan
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Correspondence to Fuh-Jyh Jan.

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Lin, CY., Ku, HM., Tsai, WS. et al. Resistance to a DNA and a RNA virus in transgenic plants by using a single chimeric transgene construct. Transgenic Res 20, 261–270 (2011). https://doi.org/10.1007/s11248-010-9412-7

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  • DOI: https://doi.org/10.1007/s11248-010-9412-7

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