Accurate identification of fungal phytopathogens is essential for virtually all aspects of plant pathology, from fundamental research on the biology of pathogens to the control of the diseases they cause. Although molecular methods, such as polymerase chain reaction (PCR), are routinely used in the diagnosis of human diseases, they are not yet widely used to detect and identify plant pathogens. Here we review some of the diagnostic tools currently used for fungal plant pathogens and describe some novel applications. Technological advances in PCR-based methods, such as real-time PCR, allow fast, accurate detection and quantification of plant pathogens and are now being applied to practical problems. Molecular methods have been used to detect several pathogens simultaneously in wheat, and to study the development of fungicide resistance in wheat pathogens. Information resulting from such work could be used to improve disease control by allowing more rational decisions to be made about the choice and use of fungicides and resistant cultivars. Molecular methods have also been applied to the study of variation in plant pathogen populations, for example detection of different mating types or virulence types. PCR-based methods can provide new tools to monitor the exposure of a crop to pathogen inoculum that are more reliable and faster than conventional methods. This information can be used to improve disease control decision making. The development and application of molecular diagnostic methods in the future is discussed and we expect that new developments will increase the adoption of these new technologies for the diagnosis and study of plant disease.

Molecular diagnostics for fungal plant pathogens

Accurate disease diagnosis and precise identification of any pathogens involved is an essential prerequisite for understanding plant diseases and controlling them effectively. Traditional methods of identifying plant pathogens can be slow and inconclusive, and this has prompted the search for alternative diagnostic techniques. Here, we review recent developments in antibody-based and nucleic acid-based diagnostic methods, and their applications in plant pathology. We also highlight some new developments in diagnostics that are likely to make an impact in the future.

Plant pathogen diagnostics : immunological and nucleic acid-based approaches

Analysis of ribosomal DNA sequences of Polymyxa species and related fungi and the development of genus- and species-specific PCR primers

Rhizomania is the most destructive disease in sugar beet throughout the world. The disease is caused by Beet necrotic yellow vein virus (BNYVV). Polymyxa betae (Keskin) is the only known vector of BNYVV, for transmission of the virus between the plants. Developing molecular diagnostic methods has a major impact on forecasting and epidemiological studies as well as screening sugar beet plants used in resistance breeding programmes. In the present study, quantum dots (QDs) were biofunctionalized with a specific antibody against P. betae. The glutathione-S-transferase protein's (GST) corresponding antibody (anti-GST) was effectively conjugated to Tioglicolicacid-modified Cadmium-Telluride QDs (CdTe-QDs) synthesized in an aqueous solution via electrostatic interaction. The dye (Rhodamine) molecules were attached to the GST. Donor–acceptor complexes (QDs-Ab-GST-Rhodamine) were then formed based on the antigen–antibody interaction. The mutual affinity of the antigen and the antibody brought the CdTeQDs...

Development of a quantum dots FRET-based biosensor for efficient detection of Polymyxa betae

Three soilborne viruses transmitted by Polymyxa betae KESKIN in sugar beet have been described: Beet necrotic yellow vein virus (BNYVV), the agent of rhizomania, Beet soilborne virus (BSBV), and Beet virus Q (BVQ). A multiplex reverse transcription-PCR technique was developed to simultaneously detect BNYVV, BSBV, and BVQ, together with their vector, P. betae. The detection threshold of the test was up to 128 times greater than that of an enzyme-linked immunosorbent assay. Systematic association of BNYVV with one or two different pomoviruses was observed. BVQ was detected in samples from Belgium, Bulgaria, France, Germany, Hungary, Italy, Sweden, and The Netherlands but not in samples from Turkey.

Multiplex Reverse Transcription-PCR for Simultaneous Detection of Beet Necrotic Yellow Vein Virus, Beet Soilborne Virus, and Beet Virus Q and Their Vector Polymyxa betae KESKIN on Sugar Beet

A sensitive DNA probe for the detection of Polymyxa betae in sugar beet roots

Development of molecular methods for the identification of Polymyxa species

The development of Diagnostics for Evaluating Beta Germplasm

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A sensitive DNA probe for the detection of Polymyxa betae in sugar beet roots

Development of molecular methods for the identification of Polymyxa species

The development of Diagnostics for Evaluating Beta Germplasm