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Real-Time PCR Detection Methods for Economically Important Grapevine Related Bacteria

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Methodologies and Results in Grapevine Research

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Abstract

Preventive measures are extremely important for control of diseases that are caused by plant pathogenic bacteria on economically important plants. For this reason, fast and reliable detection methods are required. Along with the time-consuming conventional detection methods such as isolation and culturing of bacteria on media, PCR-based methods have been introduced as supplementary tests for better diagnosis of plant pathogenic bacteria . In the case of non-culturable bacteria, such as Phytoplasma s, PCR-based tests even became indispensable. Real-time PCR has become a widely used platform in nucleic acid detection and quantification in diagnostics. Despite the general guidelines for some of the steps, there are no common guidelines or standard operating procedures for introduction of real-time PCR -based detection systems for pathogens for routine laboratory use. Four cases of detection systems for grapevine pathogenic bacteria that were developed at the National Institute of Biology are presented and discussed here, providing a practical overview of the whole process from the initial assay design to the implementation of the assay into the laboratory.

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Abbreviations

AY:

Aster yellows

BN:

Bois noir

DNA:

Deoxyribonucleic acid

ELISA:

Enzyme-linked immunosorbent assay

EPPO:

European and Mediterranean Plant Protection Organisation

FD:

Flavescence dorée

GYs:

Grapevine yellows

OEPP:

Organisation Européenne et Méditerranéenne pour la Protection des Plantes

PCR:

Polymerase chain reaction

qPCR:

Real-time PCR

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Acknowledgments

This work was partially supported by the Phytosanitary Administration and Phytosanitary Inspectorate of the Republic of Slovenia, Slovenian Ministry of Education, Science and Sport and Ministry of Agriculture, Forestry and Food. We thank Norman W. Schaad (USDA) and Igor Zidarič (Agricultural Institute of Slovenia) for bacterial strains, Gabrijel Seljak, Msc. for field work and Magda Tušek Žnidarič and Nataša Mehle for help with ELISA .

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Authors and Affiliations

  1. Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, 1000, Slovenia

    Matjaž Hren, Tanja Dreo, Jana Erjavec, Petra Nikolić, Jana Boben, Kristina Gruden, Marina Dermastia, Marjana Camloh & Maja Ravnikar

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  1. Matjaž Hren
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  2. Tanja Dreo
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  3. Jana Erjavec
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  4. Petra Nikolić
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  5. Jana Boben
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  6. Kristina Gruden
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  7. Marina Dermastia
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  8. Marjana Camloh
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  9. Maja Ravnikar
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Correspondence to Matjaž Hren .

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Editors and Affiliations

  1. , UMR Ecophysiologie et Génomique Fonction, Université de Bordeaux, Chemin de Leysotte CS 50008, Villenave d'Ornon, 33882, France

    Serge Delrot

  2. , IMEDEA-Dept. de Biologia, Universitat de les Illes Balears, Carretera Valldemossa Km. 7,5, Palma de Mallorca, 01722, Spain

    Hipólito Medrano

  3. Agriculturall Research Organisation, The Volcani Center, Institute of Horticulture, Bet Dagan, 50250, Israel

    Etti Or

  4. , Istituto di Frutti-Vitico9ltura, Universita Cattolica del Sacro Cuore, Via E. Parmense 84, Piacenza, 29100, Italy

    Luigi Bavaresco

  5. , Fondazione Edmund Mach, IASMA Research and Innovation Centre, Via E. mach 1, San Michele alll'Adige (TN), 38010, Italy

    Stella Grando

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Hren, M. et al. (2010). Real-Time PCR Detection Methods for Economically Important Grapevine Related Bacteria. In: Delrot, S., Medrano, H., Or, E., Bavaresco, L., Grando, S. (eds) Methodologies and Results in Grapevine Research. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9283-0_15

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