Autophagosome Visualization in Stressed Arabidopsis

The presence of Autophagosome in the PR was also observed by TEM. The PR zone of stressed Arabidopsis plants were excised under trinocular stereo microscope (SZ61- OLYMPUS) using sharp razor blade and fixed in 5% Glutaraldeyde in 0.1 M Cacodylate buffer (pH 7.4) 3 hours at room and transferred to 4 °C for continuation of fixation overnight. The samples were further processed according to Galsurker et al.^{55 (link)}. The final sections were placed on grids and sequentially stained with uranyl acetate and Lead citrate for 10 minutes each and viewed with Tecnai 12 TEM 100 kV (Phillips, Eindhoven, the Netherlands) equipped with MegaView II CCD camera and Analysis version −3.0 software (SoftImaging System GmbH, Münstar, Germany).

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Ambastha V., Friedmann Y, & Leshem Y. (2020). Laterals take it better – Emerging and young lateral roots survive lethal salinity longer than the primary root in Arabidopsis. Scientific Reports, 10, 3291.

Publication 2020

Tecnai 12 TEM 100 kV MegaView II CCD camera Analysis version −3.0 software

Arabidopsis Autophagosome Buffer Cacodylate Citrate Microscope Plants Uranyl acetate

Corresponding Organization :

Other organizations : Migal - Galilee Technology Center, Hebrew University of Jerusalem

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Variable analysis

independent variables

Stress applied to Arabidopsis plants

dependent variables

Presence of autophagosomes in the PR zone

control variables

Fixation time (3 hours at room temperature, continued overnight at 4°C)
Fixative (5% Glutaraldehyde in 0.1 M Cacodylate buffer, pH 7.4)
Staining (uranyl acetate and lead citrate for 10 minutes each)
Imaging equipment (Tecnai 12 TEM 100 kV (Phillips, Eindhoven, the Netherlands) equipped with MegaView II CCD camera and Analysis version −3.0 software)

controls

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