An immunofluorescence assay to detect urediniospores of Phakopsora pachyrhizi

Fulya Baysal-Gurel, Melanie L. Lewis Ivey, Anne Dorrance, Douglas Luster, Reid Frederick, Jill Czarnecki, Michael Boehm, Sally A. Miller

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

An indirect immunofluorescence spore assay (IFSA) was developed to detect urediniospores of Phakopsora pachyrhizi, 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 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.

Original languageEnglish (US)
Pages (from-to)1387-1393
Number of pages7
JournalPlant disease
Volume92
Issue number10
DOIs
StatePublished - Oct 1 2008

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Plant Science

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    Baysal-Gurel, F., Lewis Ivey, M. L., Dorrance, A., Luster, D., Frederick, R., Czarnecki, J., Boehm, M., & Miller, S. A. (2008). An immunofluorescence assay to detect urediniospores of Phakopsora pachyrhizi. Plant disease, 92(10), 1387-1393. https://doi.org/10.1094/PDIS-92-10-1387