Histochemical GUS Staining Assay

Histochemical GUS staining assay was performed using a substrate buffer: 100 mM sodium phosphate (pH 7.0), 5 mM potassium ferricyanide, 5 mM potassium ferrocyanide, 1 mM EDTA, 1% Triton-X, 1 mg/mL X-Gluc for embryo and 5 mg/mL for endosperm samples. Samples were incubated at 37 °C for 16 h and washed after the staining 3 times with 75% ethanol.

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Piskurewicz U., Sentandreu M., Iwasaki M., Glauser G, & Lopez-Molina L. (2023). The Arabidopsis endosperm is a temperature-sensing tissue that implements seed thermoinhibition through phyB. Nature Communications, 14, 1202.

Publication 2023

Assay Buffer Edta Embryo Endosperm Ethanol Potassium ferricyanide Potassium ferrocyanide Sodium phosphate

Corresponding Organization :

Other organizations : University of Geneva, University of Neuchâtel

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Variable analysis

independent variables

Substrate buffer composition (100 mM sodium phosphate (pH 7.0), 5 mM potassium ferricyanide, 5 mM potassium ferrocyanide, 1 mM EDTA, 1% Triton-X)
Concentration of X-Gluc (1 mg/mL for embryo samples, 5 mg/mL for endosperm samples)

dependent variables

GUS staining intensity in embryo and endosperm samples

control variables

Incubation temperature (37 °C)
Incubation duration (16 h)
Washing steps (3 times with 75% ethanol)

controls

No positive or negative controls were explicitly mentioned in the information provided.

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