Stability of gene silencing-based resistance to Plum pox virus in transgenic plum (Prunus domestica L.) under field conditions
Tóm tắt
Plum pox virus (PPV) is one of the most devastating diseases of Prunus species. Since few sources of resistance to PPV have been identified, transgene-based resistance offers a complementary approach to developing PPV-resistant stone fruit cultivars. C5, a transgenic clone of Prunus domestica L., containing the PPV coat protein (CP) gene, has been described as highly resistant to PPV in greenhouse tests, displaying characteristics typical of post-transcriptional gene silencing (PTGS). We show in this report that C5 trees exposed to natural aphid vectors in the field remained uninfected after 4 years while susceptible transgenic and untransformed trees developed severe symptoms within the first year. C5 trees inoculated by chip budding showed only very mild symptoms and PPV could be detected in these trees by IC-RT-PCR. The PPV-CP transgene in C5 was specifically hyper-methylated with no detectable expression. These results indicate both stability and efficiency of PTGS-based PPV resistance in plum under field conditions.
Tài liệu tham khảo
Al Rwahnih M, Myrta A, Di Terlizzi B and Boscia D (2001) First record of plum pox virus in Jordan. Acta Hor 550: 141-144.
Anandalakshmi R, Pruss G, Ge X, Marathe R, Mallory AC, Smith TH and Vance VB (1998) A viral suppressor of gene silencing in plants. Proc Natl Acad Sci USA 95: 13079-13084.
Atanassov D (1932) Plum pox. A new virus disease. Ann Univ Sofia, Fac Agric Silvic 11: 49-69.
Baulcombe DC (1996) RNA as a target and as initiator of post-transcriptional gene-silencing in transgenic plants. Plant Mol Biol 32: 79-88.
Beachy RN, Loesh-Fries S and Tumer NE (1990) Coat protein-mediated resistance against virus infection. Ann Rev Phytapathol 28: 451-474.
Brigneti G, Voinnet O, Li W-X, Ji L-H, Ding S-W and Baulcombe DC (1998) Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO 17: 6739-6746.
Dunez J and Sutic D (1988) Plum pox virus. In European Handbook of Plant Diseases (eds) Smith IM, Dunez J, Elliot RA, Phillips DH and Arches SA (pp. 44-46) Blackwell, London.
English JJ, Mueller E and Baulcombe DC (1996) Suppression of virus accumulation in transgenic plants exhibiting silencing of nuclear genes. Plant Cell 8: 179-188.
Hobbs SLA, Warkentin TD and DeLong CMO (1993) Transgene copy number can be positively or negatively associated with transgene expression. Plant Mol Biol 21: 17-26.
Ingelbrecht IL, Van Houdt H, Van Montagu M and Depicker A (1994) Post-transcriptional silencing of reporter transgenes in tobacco correlates with DNA methylation. Proc Natl Acad Sci USA 91: 10502-10506.
Jones L, Hamilton AJ, Voinnet O, Thomas CL, Maule AJ and Baulcombe DC (1999) RNA-DNA interactions and DNA methylation in post-transcriptional gene silencing. Plant Cell 12: 2291-2301.
Kegler H, Fuchs E, Gruntzig M and Shwarz S (1998) Some results of 50 years of research on the resistance to Plum pox virus. Acta Virol 42: 200-215.
Kobayashi N, Horikoshi T, Katsuyama H, Handa T and Takayanagi K (1998) A simple and efficient DNA extraction method for plants, especially woody plants. Plant Tissue Culture Biotech 4: 76-80.
Korschineck I, Himmler G, Sagl R, Steinkellner H and Katinger HWD (1991) A PCR membrane spot assay for the detection of plum pox virus RNA in bark of infected trees. J Virol Methods 31: 139-146.
Kunze L and Krczal H (1971) Transmission of sharka virus by aphids. Ann Phytopathol HS 355-260.
Levy L, Damsteegt V and Welliver R (2000) First Report of Plum PoxVirus (Sharka Disease) in Prunus persica in the United States. Plant Dis 84: 202.
Lindbo JA, Silva-Rosales L, Proebsting WM and Dougherty WG (1993) Induction of a highly specific antiviral state in transgenic plants: implications for regulation of gene expression and virus resistance. Plant Cell 5: 1749-1759.
Malinowski T, Zawadzka B, Ravelonandro M and Scorza R (1998) Preliminary report on the apparent breaking of resistance of a transgenic plum by chip bud inoculation of plum pox virus PPV-S. Acta Virol 42: 241-243.
Olmos A, Cambra M, Dasi MA, Candresse T, Esteban O, Gorris MT and Asensio M (1997) Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA. J Virol Methods 68: 127-137.
Powell-Abel P, Nelson RS, De B, Hoffman N, Rogers SG, Fraley RT and Beachy RN (1986) Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232: 738-743.
Ravelonandro M, Monsion M, Teycheney, PY, Delbos R and Dunez J (1992) Construction of a chimeric viral gene expressing plum pox virus coat protein: Gene 120: 167-173.
Ravelonandro M, Monsion M, Delbos R and Dunez J (1993) Variable resistance to plum pox virus and potato virus Y infection in transgenic Nicotiana plants expressing plum pox virus coat protein. Plant Sci 91: 157-169.
Ravelonandro M, Scorza R, Bachelier JC, Labonne G, Levy L, Damsteegt V, Callahan AM and Dunez J (1997) Resistance of transgenic Prunus domestica to plum pox virus infection. Plant Dis 81: 1231-1235.
Ravelonandro M, Dunez J, Scorza R and Labonne G (1998a) Challenging transgenic plums expressing potyvirus coat protein genes with viruliferous aphids. Acta Hort 472: 413-420.
Ravelonandro M, Scorza R, Renaud R and Salesses G (1998b) Transgenic plums resistant to plum pox virus infection and preliminary results of cross-hybridization. Acta Hort 478: 67-71.
Rosales M, Hinrichsen P and Herrera G (1998) Molecular characterization of plum pox virus isolated from apricots, plums and peaches in Chile. Acta Hort 472: 401-405.
Sanford JC and Johnston SA (1985) The concept of parasitederived resistance-deriving resistance genes from the parasite's own genome. J Theor Biol 113: 395-405.
Scorza R, Ravelonandro M, Callahan AM, Cordts JM, Fuchs M, Dunez J and Gonsalves D (1994) Transgenic plums (Prunus domestica L.) express the plum pox virus coat protein gene. Plant Cell Rep 14: 18-22.
Scorza R, Callahan AM, Levy L, Damsteegt V and Ravelonandro M (1998) Transferring potyvirus coat protein genes through hybridization of transgenic plants to produce plum pox virus resistant plums (Prunus domestica L.). Acta Hort 472: 421-427.
Scorza R, Callahan A, Levy L, Damsteegt V, Webb K and Ravelonandro M (2001) Post-transcriptional gene silencing in plum pox virus resistant transgenic European plum containing the plum potyvirus coat protein gene. Transgenic Res 10: 201-209.
Sijen T, Wellink J, Hiriart JB and van Kammen A (1996) RNA-mediated virus resistance: role of repeated transgenes and delineation of targeted regions. Plant Cell 8: 2277-2294.
Stam M, de Bruin R, Kenter S, van der Hoorn RAL, van Blokland R, Mol JNM and Kooter JM (1997) Post-transcriptional silencing of chalcone synthase in petunia by inverted transgene repeats. Plant J 12: 63-82.
Thompson D, McCann M, MacLeod M, Lye D, Green M and James D (2001) First report of plum pox potyvirus in Ontario, Canada. Plant Dis 85: 97.
Verwoerd TC, Dekker BMM and Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17: 2362.
Wilson TMA (1993) Strategy to protect crop plants against viruses: pathogen-derived resistance blossoms. Proc Natl Acad Sci USA 90: 3134-3141.