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Using Microdensitometry and Image Analysis to Study Nuclear Cycle Events in a Plant Pathogenic Fungus

Published online by Cambridge University Press:  02 July 2020

K. Snetselaar
Affiliation:
Biology Department, Saint Joseph's University, Philadelphia, PA19131
S. Lee
Affiliation:
Hitech Instruments Inc., Edgemont, PA19028
A. Nguyen
Affiliation:
Biology Department, Saint Joseph's University, Philadelphia, PA19131
M. McCann
Affiliation:
Biology Department, Saint Joseph's University, Philadelphia, PA19131
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Extract

Determining the length of the various stages of the nuclear cycle and correlating nuclear cycle events with particular aspects of cell morphology generally require the use of autoradiography. The most useful label is tritiated thymidine, because it is incorporated only into DNA. Unfortunately fungi, unlike most other organisms, do not take incorporate thymidine from medium. In the past, microspectrophotometry has been used to study fungal nuclei, but this technique measures nuclei one at a time. Recent advances in image capture and analysis software have made it possible to use the technique of photomicrodensitometry to quantify DNA in nuclei stained with fluorochromes that bind quantitatively to DNA. We are using this method to study nuclear cycle events in cells of the pathogenic fungus Ustilago maydis.

The basidiomycete fungus Ustilago maydis causes corn smut disease. This fungus is dimorphic—in addition to growing by filaments in corn plants, it can exist saprobically in a yeast-like budding stage.

Type
Light and Electron Microscopic Techniques for the Study of Plant Pathogenic Fungi and Their Interactions with Host Plants
Copyright
Copyright © Microscopy Society of America

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References

1. Muller, P. et al., Molecular Microbiology 34 (1999) 1007.CrossRefGoogle Scholar

2. Holliday, R. in Handbook of Genetics, New YorkPlenum (1974) 575.Google Scholar

3. Portions of this research were supported by NSF grant #MCB9807807 to Snetselaar, K. and McCann, M..Google Scholar