Spatial identification of transcripts and biological processes in laser micro-dissected sub-regions of waterlogged...

Over-expression of the corn phytoglobin ZmPgb1.2 increases tolerance to waterlogging, while suppression of ZmPgb1.2 compromises plant growth. To unravel compartment-specific transcriptional changes evoked by ZmPgb1.2 during hypoxia, laser micro-dissected sub-regions from waterlogged roots of WT and ZmPgb1.2 overexpressing [ZmPgb1.2(S)] plants were probed for global transcriptional analysis using next generation RNA sequencing. These sub-regions included compartments within the meristematic, elongation, and maturation zone. Of the 149 genes differentially expressed by the up-regulation of ZmPgb1.2, 78 occurred within the meristematic region and included genes involved in jasmonic acid synthesis and response, ascorbic acid metabolism, and ethylene signalling. The ZmPgb1.2 regulation of these genes, discussed in the context of known functions of Pgbs, was further validated by monitoring their expression in meristematic cells of waterlogged roots suppressing ZmPgb1.2. Of the 27 genes differentially expressed by the over-expression of ZmPgb1.2 in the elongation zone, pyruvate kinase and alcohol dehydrogenase showed an expression pattern correlated to the level of ZmPgb1.2 in the tissue. The transcriptional induction of these two enzymes in hypoxic domains of the elongation zone over-expressing ZmPgb1.2 suggests the activation of the fermentation pathway which might be required to sustain metabolic flux and production of ATP in support of cell elongation.