GCN2 and EIN2 Pathway Activation Assays in Arabidopsis Under Hypoxia

General translational repression is a crucial process that reduces energy consumption under hypoxia. Ethylene can stimulate both GCN2 and EIN2 pathways to counteract this repression and facilitate adaptation to hypoxia upon submergence. Here, we present three rapid assays to investigate the activation of GCN2 and EIN2 pathways in translation under submergence in Arabidopsis, including the survival test, eIF2α phosphorylation detection, and newly synthesized protein quantification. These methodologies will contribute to understanding ethylene signaling and plant adaptation to hypoxia stress.