An Immunofluorescence Assay to Detect Urediniospores of Phakopsora pachyrhizi

Authors

Fulya Baysal-Gurel, Ohio State University
Melanie L. Lewis Ivey, Ohio State University
Anne E. Dorrance, Ohio State UniversityFollow
Douglas Luster, USDA ARS Foreign Diseases and Weed Science Research Unit
Reid Frederick, USDA ARS Foreign Diseases and Weed Science Research UnitFollow
Jill Czarnecki, Naval Medical Research Center
Michael J. Boehm, University of Nebraska-LincolnFollow
Sally A. Miller, Ohio State University

Document Type

Article

Date of this Version

2008

Citation

Baysal-Gurel, F., Ivey, M. L. L., Dorrance, A., Luster, D., Frederick, R., Czarnecki, J., Boehm, M., and Miller, S. A. 2008. An immunofluorescence assay to detect urediniospores of Phakopsora pachyrhizi. Plant Dis. 92:1387-1393.

Comments

U.S. Government Work

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.