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Characterization of Ustilago Hordei Fimbriae Using Scanning Transmission Electron Microscopy and Immunocytochemistry

Published online by Cambridge University Press:  02 July 2020

Caroll E. Henry ,
T.L. Salaam ,
E. Steward-Clark ,
Joyce Craig  and
Lennell Reynolds
Caroll E. Henry
Affiliation:
Department of Biological Sciences, Chicago State University, Chicago, Illinois60628
T.L. Salaam
Affiliation:
Department of Biological Sciences, Chicago State University, Chicago, Illinois60628
E. Steward-Clark
Affiliation:
Department of Biological Sciences, Chicago State University, Chicago, Illinois60628
Joyce Craig
Affiliation:
Department of Biological Sciences, Chicago State University, Chicago, Illinois60628
Lennell Reynolds
Affiliation:
Department of Biological Sciences, Chicago State University, Chicago, Illinois60628
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Extract

Sporidia of Ustilago hordei produce surface fimbriae which are important in conjugation and pathogenicity. This work focuses on fimbrial origin and production using transmission electron microscopy (TEM) and immunocytochemistry.

Wild type I4A sporidial cells cultured to log phase with rotary shaking in yeast extract glucose (YEG) growth for 48 h. at 21° C, were harvested by centrifugation at 8000 rpm, placed on formvar coated grids, negatively stained with 2% uranyl acetate and photographed in the JEOL 1200 STEM. Some cells were prepared for sectioning by fixation with gluteraldehyde and cacodylate buffer, post fixed in osmium tetroxide, dehydrated and embedded in epoxy and stained with uranyl acetate. The remainder of the cells were sheared in a blender to remove fimbriae. The defimbriated cells and 1 ml. of fimbrial suspension were presented to TEM. The rest of the fimbrial suspension was centrifuged at 30,000 rpm and he fimbrial pellet protein concentration was determined to be 1.345 nm. as assayed by UV adsorption.

Type
Non-Vertebrate Biology
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 123 - 124
Copyright
Copyright © Microscopy Society of America 1997

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References

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5. The authors gratefully acknowledge the use of facilities at the Electron Microscopy Research Laboratory, Chicago State University, and the Biological Research Laboratories, University of Illinois. The work was funded by grants from NSF/RIMI and NIH/MBRS.Google Scholar