An immunofluorescence assay for the detection of wheat rust species using monoclonal antibody against urediniospores of Puccinia triticina
Article first published online: 31 JUL 2013
© 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 115, Issue 4, pages 1023–1028, October 2013
How to Cite
Gao, L., Chen, W., Liu, T. and Liu, B. (2013), An immunofluorescence assay for the detection of wheat rust species using monoclonal antibody against urediniospores of Puccinia triticina. Journal of Applied Microbiology, 115: 1023–1028. doi: 10.1111/jam.12295
- Issue published online: 16 SEP 2013
- Article first published online: 31 JUL 2013
- Accepted manuscript online: 25 JUN 2013 12:00AM EST
- Manuscript Accepted: 19 JUN 2013
- Manuscript Revised: 8 MAY 2013
- Manuscript Received: 18 FEB 2013
- National Basic Research and Development Program. Grant Numbers: 2011CB100403, 2009CB119200
- National High Technology Research and Development Program. Grant Number: 2012AA101501
- National Natural Scientific Foundation of China. Grant Number: 30800709
- Ministry of Agriculture, China. Grant Numbers: 200903035, CARS-03
- immunofluorescence assay;
- monoclonal antibody;
- plant pathology;
- Puccinia triticina ;
- wheat rust
Wheat (Triticum 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 Puccinia striiformis f. sp. tritici, Puccinia triticina and Puccinia 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.
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.