Detection, Quantification and Immunolocalisation of Botrytis species

Classical methods of detection of Botrytis species include plating-out of surface sterilized infected plant tissues, soils and airborne conidia on selective media and the identification, by microscopy, of the sclerotia, conidia and conidiophores, based on their characteristic shape, size and colour. Other methods are now available such as nucleic acid-based methods that can be used to track individual isolates or specific species. The determination of biomass levels in samples using these methods, however, is problematic because of the multinucleate nature of Botrytis conidia and thallus. Immunological methods employing genus-specific monoclonal antibodies, particularly quantitative laboratory-based plate-trapped antigen ELISAs, allow large numbers of samples to be processed easily within a few hours. These methods, combined with the modified plate spore trap, the Micro-Titre Immuno Spore Trap (MTIST), enable the quantification of conidia in microtitre wells. A rapid semiquantitative immuno-chromatographic lateral flow device designed for use in the field or office promises to be a useful screening device for Botrytis. Development of species-specific monoclonal antibodies remains a challenge. The usefulness of Fourier transform infrared spectroscopy, nuclear magnetic resonance, liquid chromatography-mass spectroscopy and enzymic methods to detect and quantify specific secondary metabolites produced by Botrytis remains to be fully demonstrated.