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ARR5::GUS assay

IS Iñigo Saiz-Fernández
MČ Martin Černý
JS Jan Skalák
BB Břetislav Brzobohatý
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An SRS was established using Arabidopsis thaliana ARR5::GUS transgenic plants, following the method described above. The plants were transferred to custom petri dishes containing separate patches of MS and MS + 10 µM transzeatin (tZ) in 0.1% DMSO. One side of the root system of each plant was placed on the medium supplemented with tZ and the other on the normal MS medium. Plants were kept for 2 days in these heterogenous growth conditions before performing the GUS staining.

The staining was performed by submerging the plants in a staining solution containing 0.1 M phosphate buffer (pH = 7.2), 0.5 mM K3[Fe(CN)6], 1% TRITON-X114, and 0.1 mg/mL X-GlcA in DMSO; and applying vacuum for 10 min. Then, the pressure was slowly restored, and plants were incubated overnight in the dark. Afterwards, the plants were washed with a destaining solution consisting of 70% (v/v) ethanol and 20% (v/v) chloroform. Pictures were taken using an Olympus IX70 microscope.

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