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An immunofluorescence assay for the detection of wheat rust species using monoclonal antibody against urediniospores of P uccinia triticina
Author(s) -
Gao L.,
Chen W.,
Liu T.,
Liu B.
Publication year - 2013
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12295
Subject(s) - urediniospore , immunofluorescence , biology , monoclonal antibody , rust (programming language) , puccinia , indirect immunofluorescence , virology , antibody , botany , microbiology and biotechnology , immunology , germination , mildew , computer science , programming language
Abstract Aims Wheat ( T riticum aestivum ) is one of the most important crop species, but yields are often drastically reduced by rust epidemics. In this report, we describe a rapid and sensitive immunofluorescence method for the detection of urediniospores of the fungi P uccinia striiformis f. sp. tritici , P uccinia triticina and P uccinia graminis f. sp. tritici , which are causal agents of wheat rust. Methods and Results The method uses monoclonal antibody LPT ‐2 against the urediniospores of P . triticina and PE ‐cy3 goat anti‐mouse. Urediniospores of P . triticina or those of two species that are difficult to distinguish from P . triticina , P . striiformis f. sp. tritici or P . graminis f. sp. tritici were immobilized on a glass slide, and the sample was then treated with LPT ‐2. Thereafter, a second antibody, goat anti‐mouse conjugated PE ‐cy3, was added, and the slide was observed in a fluoroscope. The fluorescent signal was strong with P . triticina urediniospores, weak with P . striiformis f. sp. tritici urediniospores and weak‐to‐intermediate with P . graminis f. sp. tritici urediniospores. The detection limit of this method was 2 ng ml −1 of the monoclonal antibody LPT ‐2. Conclusions In this article, we describe the production and diagnostic application of a novel mouse monoclonal antibody specific to urediniospores of P . triticina . Significance and Impact of the Study After further technical development, this method may become a tool for on‐site identification of P . triticina urediniospores and will therefore help in the selection and timing of fungicide applications for control of wheat rust outbreaks.