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Use of Cryosem and Confocal LSM to Study Pathogenesis: The Septoria Tritici Blotch Pathosystem

Published online by Cambridge University Press:  02 July 2020

Keith E. Duncan  and
Richard J. Howard
Keith E. Duncan
Affiliation:
DuPont Agricultural Products, DuPont Experimental Station, Wilmington, DE19880-0402
Richard J. Howard
Affiliation:
DuPont Agricultural Products, DuPont Experimental Station, Wilmington, DE19880-0402
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Low temperature, low voltage field emission scanning electron microscopy (cryoSEM) allows brilliant imaging of superbly preserved biological material without suffering the specimen destruction inherent in conventional SEM techniques. Confocal laser scanning microscopy (CLSM) allows observation of fluorescence-labeling within host tissues, utilizing the optical sectioning capability of the CLSM to construct composite stereo images of specimens in three dimensions. These technologies were invaluable in elucidating the septoria tritici blotch infection process on wheat, caused by Mycosphaerella graminicola (anamorph Septoria tritici).

Seedlings inoculated with M. graminicola were incubated for the first 48 h in a dew chamber with constant misting, 20°C days and 16°C nights, then transferred to a growth chamber under the same temperature conditions at 90% RH until development of mature pycnidia, visible macroscopically at 11 days post-inoculation. Samples for cryoSEM were attached to specimen blocks with Histo Prep (Fischer Scientific, Orangeburg, NY) and frozen by plunging blocks into liquid nitrogen slush.

Type
Light and Electron Microscopic Techniques for the Study of Plant Pathogenic Fungi and Their Interactions with Host Plants
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 678 - 679
Copyright
Copyright © Microscopy Society of America

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References

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