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Transgenic plums (Prunus domestica L.) express the plum pox virus coat protein gene

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Summary

Plum hypocotyl slices were transformed with the coat protein (CP) gene of plum pox virus (PPV-CP) following cocultivation with Agrobacterium tumefaciens containing the plasmid pGA482GG/PPVCP-33. This binary vector carries the PPV-CP gene construct, as well as the chimeric neomycin phosphotransferase and β-glucuronidase genes. Integration and expression of the transferred genes into regenerated plum plants was verified through kan resistance, GUS assays, and PCR amplification of the PPV-CP gene. Twenty-two transgenic clones were identified from approximately 1800 hypocotyl slices. DNA, mRNA, and protein analyses of five transgenic plants confirmed the integration of the engineered CP gene, the accumulation of CP mRNA and of PPV-CP-immunoreactive protein. CP mRNA levels ranged from high to undetectable levels, apparently correlated with gene structure, as indicated by DNA blot analysis. Western analysis showed that transgenic plants produced amounts of CP which generally correlated with amounts of detected mRNA.

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References

  • Abel PP, 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 232:738–743.

    Google Scholar 

  • Atanassov D (1932) Plum pox. A new virus disease. Annals of the University Sofia, Faculty of Agriculture and Silviculture 11:49–69.

    Google Scholar 

  • Bailey JM, Davidson N (1975) Methyl mercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem 70:75–85.

    Google Scholar 

  • Bauman G (1965) Die Ubertragung der Virosen Triebsucht auf Apfelsamlinge in Gewachshaus. Nachrichtenbl. Dt. Pflanzenschultzd. (Braunschweig) 157:73–75.

    Google Scholar 

  • Beachy RN, Loesch-Fries S, Tumer NE (1990) Coat protein-mediated resistance against virus infection. Ann Rev of Phytopath 28:451–474.

    Google Scholar 

  • Berger PH, Hunt AG, Domier LL, Hellmann GM, Stram Y, Thornbury DW, Pirone TP (1989) Expression in transgenic plants of a viral gene product that mediates insect transmission of potyvirus. Proc Natl Acad Sci USA 86:8402–8406.

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem 71:248–254.

    Google Scholar 

  • Callahan AM, Morgens PH, Wright P, Nichols Jr KE (1992) Comparison of pch313 (pTOM13 homolog) RNA accumulation during fruit softening and wounding of two phenotypically different peach cultivars. Plant Physiol 100:482–488.

    Google Scholar 

  • Caplan AB, Von Montagu M, Schell J (1985) Genetic analysis of integration mediated by single T-DNA borders. J Bact 161:655–664.

    Google Scholar 

  • Christoff A (1958) Die obstvirosen in Bulgarien. Phytopath. Z 31:381–436.

    Google Scholar 

  • Cropley R (1968) The identification of plum pox (Sharka) virus in England. Plant Path 17:66–70.

    Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Dunez J, Sutic D (1988) In European Handbook of Plant Diseases p. 44–46. Smith IM, Dunez J, Lelliot RA, Phillips DH, and Archer SA (Eds.). Blackwell Scientific Publications.

  • Fitch MMM, Manshardt RM, Gonsalves D, Slightom JL, Sanford JC (1990) Stable transformation of papaya via microprojectile bombardment. Plant Cell Reports 9:189–194.

    Google Scholar 

  • Goring DR, Thompson L, Rothstein SJ (1991) Transformation of a partial nopaline synthase into tobacco supresses the expression of a resident wild-type gene. Proc Natl Acad Sci USA 88:1770–1774

    Google Scholar 

  • Hobbs SLA, Kapodar P, DeLong CMO (1990) The effect of T-DNA copy number, position and methylation on reporter expression in tobacco transformants. Plant Mol Biol 15:851–864.

    Google Scholar 

  • Herrin DL, Schmidt FW (1988) Rapid reversible staining of northern blots prior to hybridization. Biotechniques 6:196–200

    Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep 5:387–405.

    CAS  Google Scholar 

  • Kassanis B, Sutic D (1965) Some results of recent investigations of Sharka (plum pox) virus disease. Zast. Bilja 16:335–340.

    Google Scholar 

  • Kunze L, Krczal H (1971) Transmission of Sharka virus by aphids. Eur Symp Fruit Tree Virus Dis Proc 8th:255–260.

    Google Scholar 

  • Laimer da Camara, Machado M, da Camara Machado A, Hanzer V, Weiss H, Regner F, Steinkellner H, Mattanovich D, Plail R, Knapp E, Kalthoff B, Katinger H (1992) Regeneration of transgenic plants of Prunus armeniaca containing the coat protein gene of Plum Pox Virus. Plant Cell Reports 11:25–29.

    Google Scholar 

  • Lecoq H, Ravelonandro M, Wipf-Scheibel C, Monsion M, Raccah B, Dunez J (1993) Aphid transmission of a non-aphid-transmissible strain of zucchini yellow mosaic potyvirus from transgenic plants expressing the capsid protein of plum pox potyvirus. MPMI 6:403–406

    Google Scholar 

  • Ling KS, Namba S, Gonsalves C, Slightom JL, Gonsalves D (1991) Protection against detrimental effects of Potyvirus infection in transgenic tobacco plants expressing the papaya ringspot virus coat protein gene. Bio/Technology 9:752–758.

    Google Scholar 

  • Llacer G, Cambra M, Lavina A, Arambur J (1986) Investigations on plum pox (Sharka) virus in Spain. Acta Hort 193:155–159.

    Google Scholar 

  • Louro D, Corvo LM (1986) Occurrence of Sharka in Portugal. Acta Hort 193:183–186.

    Google Scholar 

  • Mante S, Morgens PH, Scorza R, Cordts JM, Callahan AM (1991) Agrobacterium-mediated transformation of plum (Prunus domestica L.) hypocotyl slices and regeneration of transgenic plants. Bio/Technology 9:853–857.

    Google Scholar 

  • Mante S, Scorza R, Cordts JM (1989) Plant regeneration from cotyledons of Prunus persica, Prunus domestica and Prunus cerasus. Plant Cell Tis Org Cult 19:1–11.

    Google Scholar 

  • Mazyad HM, Nakhla MK, Abo-Elea A (1992) Occurrence of plum pox (Sharka) virus on stone fruit trees in Egypt. Acta Hort 309:119–124.

    Google Scholar 

  • McKinney HH (1929) Mosaic disease in the Canary Islands, West Africa, and Gibraltar. J Agric Res 39:557–558.

    Google Scholar 

  • Morgens PH, Callahan AM, Dunn L, Abeles F (1990) Isolation and sequencing of cDNA clones encoding ethylene-induced putative peroxidases from cucumber cotyledons. Plant Mol Biol 14:715–725.

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497.

    CAS  Google Scholar 

  • Nemeth J (1986) Virus, Mycoplasma, and Rickettsia Diseases of Fruit Trees. Martinus Nijhoff Publishers, Dordrecht.

    Google Scholar 

  • Ravelonandro M, Varveri C, Delbos R, Dunez J (1988) Nucleotide sequence of the capsid protein gene of plum pox potyvirus. J Gen Virol 69:1509–1516.

    Google Scholar 

  • Ravelonandro M, Monsion M, Delbos R, Dunez J (1993) Variable resistance to plum pox virus and potato virus Y injection in transgenic Nicotiana plants expressing plum pox virus coat protein. Plant Sci 91:157–169.

    Google Scholar 

  • Ravelonandro M, Monsion M, Techeney PY, Delbos R, Dunez J (1992) Construction of a chimeric viral gene allowing the expression of plum pox virus coat protein. Gene 120:167–173.

    Google Scholar 

  • Refatti E, Osler R, Loi N, Vindimian E (1985) Sharka disease in a plum area of Trento province, Italy. Riv Patol Veg 21:41–49.

    Google Scholar 

  • Regner F, da Camara Machado A, Laimer da Camara Machado M, Steinkellner H, Mattanovich D, Hanzer V, Weiss H, Katinger H (1992) Coat protein mediated resistance to Plum Pox Virus in Nicotiana clevelandii and N. benthamiana. Plant Cell Reports 11:30–33.

    Google Scholar 

  • Sanford JC, Johnston SJ (1985) The concept of parasite-derived resistance Deriving resistance genes from the parasite's own genome. J Theor Biol 113:395–405.

    Google Scholar 

  • Scorza R, Cordts JM, Mante S, Gonsalves D, Damsteegt VD, Yepes LM, Slightom JL (1991) Agrobacterium-mediated transformation of plum (Prunus domestica L.) with the coat protein gene of papaya ringspot virus. Third Int Cong Plant Mol Biol 6–11 Oct. 1991 Tuscon, AZ. abstract 1167.

  • Shaw CH, Watson MD, Carter GH, Shaw CH (1984) The right hand copy of the nopaline Ti-plasmid 25 bp repeat is required for tumour formation. Nucleic Acids Res 12:6031–6041.

    Google Scholar 

  • Sutic D, Pine TS (1968) Sharka (Plum Pox) disease. Plant Dis Reptr 52:253–256.

    Google Scholar 

  • Trifonov D (1974) Susceptibility to the plum pox virus of twelve varieties of the cultivar Kjustendilska sinya silva, Publ House Bulg Acad Sc Virology, Sofia, p. 131–138.

    Google Scholar 

  • Wang K, Herrera-Estrella L, Van Montagu M, Zambryski P (1984) Right 25 bp terminus sequence of the nopaline T-DNA is essential for and determines direction of DNA transfer from Agrobacterium to the plant genome. Cell 38:455–462.

    Google Scholar 

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Authors and Affiliations

  1. USDA-ARS Appalachian Fruit Research Station, 45 Wiltshire Rd., 25430, Kearneysville, WV, USA

    Ralph Scorza, Ann M. Callahan & John M. Cordts

  2. Station de Pathologie Vegetale, INRA, BP 81, 33 883, Villenave d'Ornon, France

    Michel Ravelonandro & Jean Dunez

  3. Department of Plant Pathology, Cornell University, 14456, Geneva, NY, USA

    Marc Fuchs & Dennis Gonsalves

Authors
  1. Ralph Scorza
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  2. Michel Ravelonandro
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  3. Ann M. Callahan
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  4. John M. Cordts
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  5. Marc Fuchs
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  6. Jean Dunez
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  7. Dennis Gonsalves
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Communicated by R. N. Beachy

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Scorza, R., Ravelonandro, M., Callahan, A.M. et al. Transgenic plums (Prunus domestica L.) express the plum pox virus coat protein gene. Plant Cell Reports 14, 18–22 (1994). https://doi.org/10.1007/BF00233291

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  • DOI: https://doi.org/10.1007/BF00233291

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