An immunofluorescence assay to detect soybean rust urediniospores

Authors: BAYSAL, FULYA - OHIO STATE UNIVERSITY; DORRANCE, ANNE - OHIO STATE UNIVERSITY; LEWIS-IVEY, MELANIE - OHIO STATE UNIVERSITY; Luster, Douglas - Doug; Frederick, Reid; CZARNECKI, JILL - US NAVY BIO DEF RESEARCH; BOEHM, MICHAEL - OHIO STATE UNIVERSITY; MILLER, SALLY - OHIO STATE UNIVERSITY

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/2007
Publication Date: 8/1/2007
Citation: Baysal, F., Dorrance, A., Lewis-Ivey, M., Luster, D.G., Frederick, R.D., Czarnecki, J., Boehm, M., Miller, S. 2007. An immunofluorescence assay to detect soybean rust urediniospores. Phytopathology. 97:S9

Interpretive Summary:

Technical Abstract: An indirect immunofluorescence assay (IF) was developed to detect urediniospores of Phakopsora pachyrhizi and P. meibomiae, utilizing rabbit polyclonal antisera produced in response to intact non-germinated (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 assays. SBR1A and SBR2 binding to spores was detected with goat anti-rabbit Alexa Fluor 488-tagged antiserum using a Leica DM IRB epifluorescent microscope with an I3 blue filter, (excitation 450-490 nm, emission 515 nm). Both antisera bound to spores of P. pachyrhizi and P. meibomiae, although SBR2 bound P. meibomiae spores more weakly than SBR1A. The assay was carried out in solution in test tubes and on standard glass microscope slides. Double-stick tape affixed to slides was superior to petroleum jelly both in retaining spores and in immunofluorescence due to non-specific binding of the Alexa Fluor 488-tagged antisera to the latter. The assay was used to confirm the identity of P. pachyrhizi urediniospores captured on glass slides in passive air samplers in Georgia, Kentucky and Ohio during 2006.