TUNEL Assay and DAPI Staining Revealed Few Alterations of Cellular Morphology in Naturally and Artificially Aged Seeds of Cultivated Flax

In a search for useful seed aging signals as biomarkers for seed viability prediction, we conducted an experiment using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and 4′,6-diamidino-2-phenylindole (DAPI) staining to analyze morphological and molecular changes in naturally aged (NA) and artificially aged (AA) flax (Linum usitatissimum L.) seeds. A total of 2546 sections were performed from 112 seeds of 12 NA and AA seed samples with variable germination rates. Analyzing 1384 micrographs generated from TUNEL assay and DAPI staining revealed few alterations of the cellular morphology of the NA and AA seeds. Also, the revealed DNA degradations in the aged flax seeds appeared to be associated with seed samples of low germination rates. These results suggest that oily flax seed aging may alter the cellular morphology differently than starchy wheat seed aging. The results also imply that the TUNEL assay and DAPI staining may not yield informative assessments on cellular alterations and DNA degradation after the aging of oily seeds.