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An Immunofluorescence Assay to Detect Urediniospores of Phakopsora pachyrhizi

October 2008 , Volume 92 , Number  10
Pages  1,387 - 1,393

Fulya Baysal-Gurel , Melanie L. Lewis Ivey , and Anne Dorrance , Department of Plant Pathology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster ; Douglas Luster and Reid Frederick , USDA ARS Foreign Diseases and Weed Science Research Unit, Ft. Detrick, MD ; Jill Czarnecki , Naval Medical Research Center, Biological Defense Research Directorate, Silver Spring, MD ; Michael Boehm , Department of Plant Pathology, The Ohio State University, Columbus ; and Sally A. Miller , Department of Plant Pathology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster



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Accepted for publication 6 June 2008.
ABSTRACT

An indirect immunofluorescence spore assay (IFSA) was developed to detect urediniospores of Phakopsora pachyrhizi, utilizing rabbit polyclonal antisera produced in response to intact nongerminated (SBR1A) or germinated (SBR2) urediniospores of P. pachyrhizi. Both antisera were specific to Phakopsora spp. and did not react with other common soybean pathogens or healthy soybean leaf tissue in enzyme-linked immunosorbent assay (ELISA). SBR1A and SBR2 bound to P. pachyrhizi and P. meibomiae urediniospores were detected with goat anti-rabbit Alexa Fluor 488-tagged antiserum using a Leica DM IRB epifluorescent microscope with an I3 blue filter (excitation 450 to 490 nm, emission 515 nm). The assay was performed on standard glass microscope slides; double-sided tape was superior to a thin coating of petroleum jelly both in retaining spores and in immunofluorescence. The IFSA was used to confirm the identity of P. pachyrhizi urediniospores captured on glass slides from passive air samplers from Georgia, Kentucky, and Ohio during 2006.


Additional keywords: field trap, soybean rust

© 2008 The American Phytopathological Society