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International Microbiology

, Volume 6, Issue 4, pp 233–243 | Cite as

Innovative tools for detection of plant pathogenic viruses and bacteria

  • María M. LópezEmail author
  • Edson Bertolini
  • Antonio Olmos
  • Paola Caruso
  • María Teresa Gorris
  • Pablo Llop
  • Ramón Penyalver
  • Mariano Cambra
Review Article

Abstract

Detection of harmful viruses and bacteria in plant material, vectors or natural reservoirs is essential to ensure safe and sustainable agriculture. The techniques available have evolved significantly in the last few years to achieve rapid and reliable detection of pathogens, extraction of the target from the sample being important for optimising detection. For viruses, sample preparation has been simplified by imprinting or squashing plant material or insect vectors onto membranes. To improve the sensitivity of techniques for bacterial detection, a prior enrichment step in liquid or solid medium is advised. Serological and molecular techniques are currently the most appropriate when high numbers of samples need to be analysed. Specific monoclonal and/or recombinant antibodies are available for many plant pathogens and have contributed to the specificity of serological detection. Molecular detection can be optimised through the automatic purification of nucleic acids from pathogens by columns or robotics. New variants of PCR, such as simple or multiplex nested PCR in a single closed tube, co-operative-PCR and real-time monitoring of amplicons or quantitative PCR, allow high sensitivity in the detection of one or several pathogens in a single assay. The latest development in the analysis of nucleic acids is micro-array technology, but it requires generic DNA/RNA extraction and pre-amplification methods to increase detection sensitivity. The advances in research that will result from the sequencing of many plant pathogen genomes, especially now in the era of proteomics, represent a new source of information for the future development of sensitive and specific detection techniques for these microorganisms.

Keywords

Antibodies Co-operational PCR DNA microarrays ELISA Enrichment FISH Multiplex PCR Nested-multiplex PCR Real time PCR 

Notes

Acknowledgements

The authors are grateful to all their colleagues at IVIA who contributed to the development of different techniques. We wish to thank the project QLK5-CT 2001-02270 granted by EU and the projects AGL 2000-0394-P4-04 and CAO 00-007 financed by the Spanish Ministerio de Ciencia y Tecnología and the Spanish Ministerio de Agricultura, Pesca y Alimentacion, respectively.

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Copyright information

© Springer-Verlag and SEM 2003

Authors and Affiliations

  • María M. López
    • 1
    Email author
  • Edson Bertolini
    • 1
  • Antonio Olmos
    • 1
  • Paola Caruso
    • 1
  • María Teresa Gorris
    • 1
  • Pablo Llop
    • 1
  • Ramón Penyalver
    • 1
  • Mariano Cambra
    • 1
  1. 1.Instituto Valenciano de Investigaciones AgrariasValenciaSpain

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