Plant Cell Reports

, Volume 27, Issue 1, pp 57–66 | Cite as

Genetic transformation of sweet orange with the coat protein gene of Citrus psorosis virus and evaluation of resistance against the virus

  • María Cecilia ZanekEmail author
  • Carina Andrea Reyes
  • Magdalena Cervera
  • Eduardo José Peña
  • Karelia Velázquez
  • Norma Costa
  • Maria Inés Plata
  • Oscar Grau
  • Leandro Peña
  • María Laura García
Genetic Transformation and Hybridization


Citrus psorosis is a serious viral disease affecting citrus trees in many countries. Its causal agent is Citrus psorosis virus (CPsV), the type member of genus Ophiovirus. CPsV infects most important citrus varieties, including oranges, mandarins and grapefruits, as well as hybrids and citrus relatives used as rootstocks. Certification programs have not been sufficient to control the disease and no sources of natural resistance have been found. Pathogen-derived resistance (PDR) can provide an efficient alternative to control viral diseases in their hosts. For this purpose, we have produced 21 independent lines of sweet orange expressing the coat protein gene of CPsV and five of them were challenged with the homologous CPV 4 isolate. Two different viral loads were evaluated to challenge the transgenic plants, but so far, no resistance or tolerance has been found in any line after 1 year of observations. In contrast, after inoculation all lines showed characteristic symptoms of psorosis in the greenhouse. The transgenic lines expressed low and variable amounts of the cp gene and no correlation was found between copy number and transgene expression. One line contained three copies of the cp gene, expressed low amounts of the mRNA and no coat protein. The ORF was cytosine methylated suggesting a PTGS mechanism, although the transformant failed to protect against the viral load used. Possible causes for the failed protection against the CPsV are discussed.


Citrus psorosis virus PDR Post-transcriptional gene silencing Citrus Transgenic trees 



Citrus psorosis virus


Coat protein


Triple sandwich immunoassay


Pathogen-derived resistance


Cauliflower mosaic virus


Alfalfa mosaic virus


Nopaline synthase gene


Optical density


Days post inoculation


Infected control


Non-infected control


Post-transcriptional gene silencing


Citrus tristeza virus


Grapevine fan-leaf virus


Tomato spotted wilt virus



We thank Dr. L.W. Timmer for helpful discussion and manuscript revision, and Ing. G. Chiarrone and Pto. Agr. Fabián Ramos for greenhouse work. M. L. García, Eduardo J. Peña, Carina A. Reyes and M. C. Zanek belong to the staff of Facultad de Ciencias Exactas, UNLP, O.G. is recipient of the research career award from CICBA, and MLG from CONICET. Lic. Zanek and Reyes are fellows of CONICET. Lic. E.J. Peña is fellows of ANPCyT. This work was supported by grants from BID802 OC-AR PICT 6198 SECyT-CONICET, AECI, CICBA and INTA EEA Concordia.


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • María Cecilia Zanek
    • 1
    Email author
  • Carina Andrea Reyes
    • 1
  • Magdalena Cervera
    • 3
  • Eduardo José Peña
    • 1
  • Karelia Velázquez
    • 1
  • Norma Costa
    • 2
  • Maria Inés Plata
    • 2
  • Oscar Grau
    • 1
  • Leandro Peña
    • 3
  • María Laura García
    • 1
  1. 1.Facultad de Ciencias ExactasInstituto de Bioquímica y Biología Molecular (IBBM), U.N.L.P.La PlataArgentina
  2. 2.Estación Experimental Agropecuaria, INTAConcordiaArgentina
  3. 3.Dpto de Protección Vegetal y BiotecnologiaInstituto Valenciano de Investigaciones Agrarias (IVIA)MoncadaSpain

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