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No recombination detected in artificial potyvirus mixed infections and between potyvirus derived transgenes and heterologous challenging potyviruses

Published online by Cambridge University Press:  16 November 2007

Christof Dietrich ,
Jane Miller ,
Gaynor Mckenzie ,
László Palkovics ,
Ervin Balázs ,
Peter Palukaitis  and
Edgar Maiss
Christof Dietrich
Affiliation:
German Collection of Microorganisms and Cell Cultures, Plant Virus Division, Inhoffenstraße 7b, 38124 Braunschweig, Germany
Jane Miller
Affiliation:
Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, Scotland, UK
Gaynor Mckenzie
Affiliation:
Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, Scotland, UK
László Palkovics
Affiliation:
Department of Plant Pathology, Corvinus University Budapest, 1118 Villányi ut 29, Hungary
Ervin Balázs
Affiliation:
Department of Applied Genomics, Agricultural Research Institute, 2462 Martonvásár, Brunszvik u 2, Hungary
Peter Palukaitis
Affiliation:
Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, Scotland, UK
Edgar Maiss
Affiliation:
Institute of Plant Diseases and Plant Protection, University of Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany

Abstract

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Risk-assessment studies of virus-resistant transgenic plants (VRTPs) focussing on recombination of a plant virus with a transgenic sequence of a different virus should include a comparison of recombination frequencies between viruses in double-infected non-transgenic plants with those observed in singly infected transgenic plants to estimate recombination incidence in VRTPs. In this study, the occurrence of recombination events was investigated in non-transgenic plants double-infected with two different potyviruses, as well as in potyviral genomes in singly infected transgenic plants expressing potyvirus sequences. Different potyviruses, namely Potato virus A (PVA), Tobacco vein mottling virus (TVMV), two strains of Potato virus Y (PVY-O, PVY-H) and two strains of Plum pox virus (PPV-NAT, PPV-SK68), were used in three combinations for double infection of a common host. Furthermore, transgenic plants expressing either potyviral coat protein (CP), helicase (CI) or polymerase (NIb) coding sequences (PPV-NAT-CP, PVY-CI, PVY-NIb) were singly-infected with a heterologous potyvirus, which was not targeted by the respective transgenic resistance. To identify recombinant potyviral sequences, a sensitive RT-PCR was developed to detect up to one recombinant molecule out of 106 parental molecules. In 304 mixed infected non-transgenic plants, 92 mixed and 164 single infected transgenic plants screened for recombinant sequences no recombinant potyviral sequence was found. These results indicate that recombination events between different potyviruses in mixed infections and between a potyvirus infecting a potyvirus-resistant transgenic plant are likely to be very infrequent.

Keywords

potyvirustransgenic plantsmixed infectionsrecombinationRT-PCR
Type
Research Article
Information
Environmental Biosafety Research , Volume 6 , Issue 3 , July 2007 , pp. 207 - 218
Copyright
© ISBR, EDP Sciences, 2007

References

Aaziz, R, Tepfer, M (1999a) Recombination in RNA viruses and in virus-resistent transgenic plants. J. Gen. Virol. 80: 1339–1346 CrossRef
Aaziz, R, Tepfer, M (1999b) Recombination between genomic RNAs of two cucumoviruses under conditions of minimal selection pressure. Virology 263: 282–289 CrossRef
Audy, P, Palukaitis, P, Slack, SA, Zaitlin, M (1994) Replicase-mediated resistance to potato virus Y in transgenic tobacco plants. Mol. Plant-Microbe Interact. 7: 15–22 CrossRef
Beachy, RN (1997) Mechanisms and applications of pathogen-derived resistance in transgenic plants. Curr. Opin. Biotechnol. 8: 215–220 CrossRef
Boonham, N, Walsh, K, Preston, S, North, J, Smith, P, Barker I (2002) The detection of tuber necrotic isolates of Potato virus Y, and the accurate discrimination of PVY(O), PVY(N) and PVY(C) strains using RT-PCR. J. Virol. Methods 102: 103–112 CrossRef
Borja, M, Rubio, T, Scholthof, HB, Jackson, AO (1999) Restoration of wild-type virus by double recombination of tombusvirus mutants with a host transgene. Mol. Plant-Microbe Interact. 12: 153–162 CrossRef
Bousalem, M, Dallot, S, Fuji, S, Natsuaki, KT (2003) Origin, world-wide dispersion, bio-geographical diversification, radiation and recombination: an evolutionary history of Yam mild mosaic virus (YMMV). Infect. Genet. Evol. 3: 189–206 CrossRef
Bujarski, JJ, Nagy, PD (1996) Different mechanisms of homologous and nonhomologous recombination in Brome mosaic virus: role of RNA sequences and replicase proteins. Semin. Virol. 7: 363–372 CrossRef
Cervera, MT, Riechmann, JL, Martín, MT, García, JA (1993) $3'$ -Terminal sequence of Plum pox virus PS and o6 isolates: evidence for RNA recombination within the potyvirus group. J. Gen. Virol. 74: 329–334 CrossRef
de Wispelaere, M, Gaubert, S, Trouilloud, S, Belin, C, Tepfer, M (2005) A map of the diversity of RNA3 recombinants appearing in plants infected with cucumber mosaic virus and tomato asperny virus. Virology 331: 117–127 CrossRef
Dietrich, C, Maiss, E (2002) Red fluorescent protein DsRed from Discosoma sp. as a reporter protein in higher plants. Biotechniques 32: 286–293
Dietrich, C, Maiss, E (2003) Fluorescent labelling reveals spatial separation of potyvirus populations in mixed infected Nicotiana benthamiana plants. J. Gen. Virol. 84: 2871–2876 CrossRef
Domier, LL, Franklin, KM, Shahabuddin, M, Hellmann, GM, Overmeyer, JH, Hiremath, ST, Siaw, MFE, Lomonossoff, GP, Shaw, JG, Rhoads, RE (1986) The nucleotide sequence of Tobacco vein mottling virus RNA. Nucleic Acids Res. 14: 5417–5430 CrossRef
Fernandez-Cuartero, B, Burgyan, J, Aranda, MA, Salanki, K, Moriones, E, Garcia-Arenal F (1994) Increase in the relative fitness of a plant virus RNA associated with its recombinant nature. Virology 203: 373–377 CrossRef
Fernandez-Delmond, I, Pierrugues, O, de Wispelaere, M, Guilbaud, L, Gaubert, S, Diveki, Z, Godon, C, Tepfer, M, Jacquemond, M (2004) A novel strategy for creating recombinant infectious RNA virus genomes. J. Virol. Methods 121: 247–257 CrossRef
Gal-On, A, Meiri, E, Raccah, B, Gaba, V (1998) Recombination of engineered defective RNA species produces infective potyvirus in planta. J. Virol. 72: 5268–5270
Gibbs, MJ (1994) Risks in using transgenic plants? Science 264: 1650–1651 CrossRef
Glais, L, Tribodet, M, Kerlan, C (2002) Genomic variability in Potato potyvirus Y (PVY): evidence that PVY(N)W and PVY(NTN) variants are single to multiple recombinants between PVY(O) and PVY(N) isolates. Arch. Virol. 147: 363–378 CrossRef
Glasa, M, Palkovics, L, Kominek, P, Labonne, G, Pittnerova, S, Kudela, O, Candresse, T, Subr, Z (2004) Geographically and temporally distant natural recombinant isolates of Plum pox virus (PPV) are genetically very similar and form a unique PPV subgroup. J. Gen. Virol. 85: 2671–2681 CrossRef
Glasa, M, Marie-Jeann, V, Moury, B, Kúdela, O, Quiot, J-B (2002) Molecular variability of the P3-6K1 region among geographically and biologically distinct isolates of Plum pox virus. Arch. Virol. 147: 563–575 CrossRef
Greene, AE, Allison, RF (1994) Recombination between viral-RNA and transgenic plant transcripts. Science 263: 1423–1425 CrossRef
Greene, AE, Allison, RF (1996) Deletions in the 3' untranslated region of cowpea chlorotic mottle virus transgene reduce recovery of recombinant viruses in transgenic plants. Virology 225: 231–234 CrossRef
Jridi, C, Martin, JF, Marie-Jeanne, V, Labonne, G, Blanc, S (2006) Distinct viral populations differentiate and evolve independently in a single perennial host plant. J. Virol. 80: 2349–2357 CrossRef
Koenig, R, Büttner, G (2004) Strategies for the detection of potential Beet necrotic yellow vein virus genome recombinations which might arise as a result of growing A type coat protein gene-expressing sugarbeets in soil containing B type virus. Transgenic Res. 13: 21–28 CrossRef
Korte, AM, Maiss, E, Kramer, I, Casper, R (1995) Biosafety considerations of different Plum pox potyvirus (PPV) genes used for transformation of plants. Acta Hort. 368: 280–284 CrossRef
Krause-Sakate, R, Fakhfakh, H, Peypelut, M, Pavan, MA, Zerbini, FM, Marrakchi, M, Candresse, T, Le Gall O (2004) A naturally occurring recombinant isolate of Lettuce mosaic virus. Arch. Virol. 149: 191–197
Lawson, C, Kaniewski, W, Haley, L, Rozman, R, Newell, C, Sanders, P, Tumer, NE (1990) Engineering resistance to mixed virus infection in a commercial potato cultivar: resistance to Potato virus X and Potato virus Y in transgenic Russet Burbank. Biotechnology 8: 127–134
López-Moya, JJ, Fernández-Ferández, MR, Cambra, M, García, JA (2000) Biotechnological aspects of Plum pox virus. J. Biotechnol. 76: 121–136 CrossRef
Maiss, E, Timpe, U, Brisske-Rode, A, Lesemann, D-E, Casper R (1992) Infectious in vivo transcripts of a Plum pox potyvirus full-length cDNA clone containing the Cauliflower mosaic virus 35S RNA promoter. J. Gen. Virol. 73: 709–713 CrossRef
Meier, M, Truve, E (2006) An attempt to identify recombinants between two sobemoviruses in doubly infected oat plants. Environ. Biosafety Res. 5: 47–56 CrossRef
Menzel, W, Jelkmann, W, Maiss, E (2002) Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control. J. Virol. Meth. 99: 81–92 CrossRef
Moonan, F, Molina, J, Mirkov, TE (2000) Sugarcane yellow leaf virus: an emerging virus that has evolved by recombination between luteoviral and poleroviral ancestors. Virology 269: 156–171 CrossRef
Moreno, IM, Malpica, JM, Diaz-Pendon, JA, Moriones, E, Fraile, A, García-Arenal F (2004) Variability and genetic structure of the population of Watermelon mosaic virus infecting melon in Spain. Virology 318: 451–60 CrossRef
Moreno, M, Bernal, JJ, Jimenez, I, Rodriguez-Cerezo E (1998) Resistance in plants transformed with the P1 or P3 gene of Tobacco vein mottling potyvirus. J. Gen. Virol. 79: 2819–2827 CrossRef
Palkovics, L, Burgyan, J, Balázs, E (1993) Comparative sequence analysis of four complete primary structures of Plum pox virus strains. Virus Genes 7: 339–347 CrossRef
Powell-Abel, P, Nelson, RS, De, B, Hoffmann, N, Rogers, SG, Fraley, RT, Beachy, RN (1986) Delay of disease development in transgenic plants that express the Tobacco mosaic virus coat protein gene. Science 232: 738–763 CrossRef
Revers, F, Le Gall, O, Candresse, T, Le Romancer, M, Dunez J (1996) Frequent occurrence of recombinant potyvirus isolates. J. Gen. Virol. 77: 1953–1965 CrossRef
Rubio, T, Borja, M, Scholthoff, HB, Jackson, AO (1999) Recombination with host transgenes and effect on virus evolution: an overview and opinion. Mol. Plant-Microbe Interact. 12: 87–92 CrossRef
Savenkov, EI, Valkonen, JPT (2001) Coat protein gene-mediated resistance to Potato virus A in transgenic plants is suppressed following infection with another potyvirus. J. Gen. Virol. 82: 2275–2278 CrossRef
Shukla, DD, Ward, CW (1988) Amino acid sequence homology of coat protein as a basis for identification and classification of the potyvirus group. J. Gen. Virol. 69: 2703–2710 CrossRef
Slilaty, SN, Lebel, S (1998) Accurate insertional inactivation of lacZ $\alpha$ : construction of pTrueBlue and M13TrueBlue cloning vectors. Gene 213: 83–91 CrossRef
Tan, Z, Wada, Y, Chen, J, Ohshima, K (2004) Inter- and intralineage recombinants are common in natural populations of Turnip mosaic virus. J. Gen. Virol. 85: 2683–2696 CrossRef
Tepfer, M (2002) Risk assessment of virus-resistant transgenic plants. Annu. Rev. Phytopathol. 40: 467–491 CrossRef
Thole, V, Dalmay, T, Burgyan, J, Balázs, E (1993) Cloning and sequencing of potato virus Y (Hungarian isolate) genomic RNA. Gene 123: 149–156 CrossRef
Urcuqui-Inchima, S, Haenni, AL, Bernardi, F (2001) Potyvirus proteins: a wealth of functions. Virus Res. 74: 157–175 CrossRef
Varrelmann, M, Maiss, E (2000) Mutations in the coat protein gene of Plum pox virus suppress particle assembly, heterologous encapsidation and complementation in plants of Nicotiana benthamiana. J. Gen. Virol. 81: 567–576 CrossRef
Varrelmann, M, Palkovics, L, Maiss, E (2000) Transgenic or plant expression vector-mediated recombination of Plum pox virus. J. Virol. 74: 7462–7469 CrossRef
Wintermantel, WM, Schoelz, JE (1996) Isolation of recombinant viruses between Cauliflower mosaic virus and a viral gene in transgenic plants under conditions of moderate selection pressure. Virology 223: 156–164 CrossRef