Identification of hypoxia-inducible genes in Arabidopsis mesophyll cells

The goal of this experiment was to investigate the early transcript changes (6h) induced by hypoxia treatment in mesophyll protoplasts. A single pair (control & hypoxia) of GeneChips® was used to confirm that hypoxia treatment altered the expression of an overlapping set of genes controlled by KIN10 (At3g01090) in Arabidopsis mesophyll protoplasts. Keywords: KIN10, KIN11, darkness, hypoxia, starvation, stress, sugar signalling, Arabidopsis, SnRK1 For the hypoxic treatment protoplasts were submerged in mannitol buffer and the hypoxia condition was confirmed by the activation of the well-established early marker genes ADH (alcohol dehydrogenase, At1g77120) and PDC1 (pyruvate decarboxylase-1, At4g33070) based on RT-PCR and GeneChip® analyses. Results show a clear overlap in genes induced under hypoxic conditions and those induced by KIN10 expression in protoplasts. The overlap of the gene expression profiles was confirmed by triplicated real-time quantitative RT-PCR analyses of selected marker genes.