Skip to main content
SpringerLink
Log in

Indexing of Citrus Viroids by Imprint Hybridisation

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

A method based on the hybridisation of tissue imprints was developed for routine indexing of citrus viroids. For maximum sensitivity and reliability, the inoculation of Citrus medica (Etrog citron) as a viroid amplification host is required. Hybridisation against Digoxigenin-labelled RNA- or DNA-probes followed by detection of viroid-probe hybrids using anti-DIG-alkaline phosphate conjugate and the chemiluminescence substrate CSPD was suitable for the detection of all citrus viroids with the same sensitivity as other available methods. The overall process is extremely simple and allows quick analysis of large numbers of samples by easily trained personnel and minimum equipment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Astruc N, Marcos JF, Macquaire G, Candresse T and Pallás V (1996) Studies on the diagnosis of hop stunt viroid in fruit trees: identification of new hosts and application of a nucleic acid extraction procedure based on non-organic solvents. European Journal of Plant Pathology 102: 837-846

    Google Scholar 

  • Davino M, Pelicani L, Renis M and Albanese G (1991) Homology of hop stunt viroid with citrus cachexia viroid. In: Proceedings of the 11th Conference of the International Organization of Citrus Virologists (IOCV), Riverside, pp. 196-201

  • Duran-Vila N, Pina JA and Navarro L (1991) Exclusion and/or uneven distribution of viroids in four citrus hosts. In: Proceedings of the 11th Conference of the International Organization of Citrus Virologists (IOCV), Riverside, pp. 219-223

  • Duran-Vila N, Pina JA and Navarro L (1993) Improved indexing of citrus viroids. In: Proceedings of the 12th Conference of the International Organization of Citrus Virologists (IOCV), Riverside, pp. 202-211

  • Duran-Vila N, Romero-Durbán J and Hernández M (1996) Detection and eradication of chrysanthemum stunt in Spain. OEPP Bulletin 26: 399-405

    Google Scholar 

  • Duran-Vila N, Roistacher CN, Rivera-Bustamante R and Semancik JS (1988) A definition of citrus viroids groups and their relationship to the exocortis disease. Journal of General Virology 69: 3069-3080

    Google Scholar 

  • Flores R (1988) Detection of citrus exocortis viroid in natural and experimental citrus hosts by biochemical methods. In: Proceedings of the 10th Conference of the International Organization of Citrus Virologists (IOCV), Riverside, pp. 192-196

  • Flores R, Randles JW, Bar-Joseph M and Diener TO (1998) A proposed scheme for viroid classification and nomenclature. Arch Virol 143: 623-629

    Google Scholar 

  • Gross HJ, Krupp G, Domdey H, Raba M, Jank P, Lossow C, Alberty H, Ramm K and Sänger HL (1982) Nucleotide sequence and secondary structure of citrus exocortis and chrysanthemum stunt viroid. European Journal of Biochemistry 121: 249-257

    Google Scholar 

  • Hadas R, Bar-Joseph M and Semancik JS (1989) Segregation of a viroid complex from a graft-transmissible dwarfing agent source from grapefruit trees. Annals of Applied Biology 115: 515-520

    Google Scholar 

  • La Rosa R, Albanese G, Azzaro A, Sesto F and Domina F (1988) In: Proceedings of the 10th Conference of the International Organization of Citrus Virologists (IOCV), Riverside, pp. 188-191

  • Maniatis T, Fritsh EF and Sambrook J (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Ohno T, Takamatsu N, Meshi T and Okada Y (1983) Hop stunt viroid: molecular cloning and nucleotide sequence of the complete cDNA copy. Nucleic Acids Research 11: 6158-6197

    Google Scholar 

  • Owens RA and Diener TO (1981) Sensitive and rapid diagnosis of potato spindle tuber viroid disease by nucleic acid hybridization. Science 213: 670-672

    Google Scholar 

  • Podleckis EV, Hammod RW, Hurtt SS and Hadidi A (1993) Chemiluminescent detection of potato and pome fruit viroids by digoxigenin-labelled dot-blot and tissue-blot hybridization. Journal of Virological Methods 43: 147-158

    Google Scholar 

  • Puchta H, Ramm K, Luckinger R, Hadas R, Bar-Joseph M and Sänger HL (1991) Primary and secondary structure of citrus viroid IV (CVd IV), a new chimeric viroid present in dwarfed grapefruit in Israel. Nucleic Acids Research 19: 6640

    Google Scholar 

  • Rakowski AG, Szychowski JA, Avena ZS and Semancik JS (1994) Nucleotide sequence and structural features of the Group III citrus viroids. Journal of General Virology 75: 3581-3584

    Google Scholar 

  • Roistacher CN, Bash JA and Semancik JS (1993) Distinct disease symptoms in Poncirus trifoliata induced by three citrus viroids from three specific groups. In: Proceedings of the 12th Conference of the International Organization of Citrus Virologists (IOCV), Riverside, pp. 173-179

  • Roistacher CN, Blue RL and Calavan EC (1973) A new test for citrus cachexia. Citrograph 58: 261-262

    Google Scholar 

  • Roistacher CN, Calavan EC, Blue RL, Navarro L and Gonzales R (1977) A new more sensitive citron indicator for the detection of mild isolates of citrus exocortis viroid (CEV). Plant Disease Reporter 61: 135-139

    Google Scholar 

  • Romero-Durbán J, Cambra M and Duran-Vila N (1995) A simple imprint-hybridization method for detection of viroids. Journal of Virological Methods 55: 37-47

    Google Scholar 

  • Sano T, Kudo H, Sugimoto T and Shikata E (1988) Synthetic oligonucleotide hybridization probes to diagnose hop stunt viroid strains and citrus exocortis viroid. Journal of Virological Methods 19: 109-119

    Google Scholar 

  • Semancik JS and Weathers LG (1972) Exocortis disease: evidence for a new species of ‘infectious’ low molecular weight RNA in plants. Nature New Biology 237: 242-244

    Google Scholar 

  • Semancik JS, Rakowski AG, Bash JA and Gumpf DJ (1997) Application of selected viroids for dwarfing and enhancement of production of ‘Valencia’ orange. Journal of Horticultural Science 72: 563-570

    Google Scholar 

  • Semancik JS, Roistacher CN, Rivera-Bustamante R and Duran-Vila N (1988) Citrus cachexia viroid, a new viroid of citrus: relationship to viroids of the exocortis disease complex. Journal of General Virology 69: 3059-3068

    Google Scholar 

  • Visvader JE and Symons RH (1985) Eleven new sequence variants of citrus exocortis viroid and the correlation of sequence with pathogenicity. Nucleic Acids Research 13: 2907-2920

    Google Scholar 

  • Yang X, Hadidi A and Garnsey SM (1992) Enzymatic cDNA amplification of citrus exocortis and cachexia viroids from infected citrus hosts. Phytopathology 82: 279-285

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias, Apartado Oficial, 46113, Moncada (Valencia), Spain

    A. Palacio-Bielsa, X. Foissac & N. Duran-Vila

Authors
  1. A. Palacio-Bielsa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. X. Foissac
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N. Duran-Vila
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to N. Duran-Vila.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Palacio-Bielsa, A., Foissac, X. & Duran-Vila, N. Indexing of Citrus Viroids by Imprint Hybridisation. European Journal of Plant Pathology 105, 897–903 (1999). https://doi.org/10.1023/A:1008711402856

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1008711402856

Search

Navigation