Characterization of cold temperature mutants

Four replicates of ~50 seeds from several overexpressing mutants of Arabidopsis (Arabidopsisthaliana (L.) Heynh.), that complete germination faster than the Columbia wild type (Col), were surface sterilized along with wild-type control seeds, and sown on 1% (w/v) agarose solid media in a Petri dish. Each of the four 60 X 15 mm Petri dishes was bifurcated by a line and, in each of the four plates, one replicate of 50 wild-type seeds was sown on one side of the line and 50 mutant seeds were sown on the other side. The plates were sealed and the four dishes were arranged side by side on the flatbed scanner inside a germinator maintaining a constant temperature at 10°C ± 1°C and constant light (135 |lmol/m2/s). Images were captured every hour starting on the third day of the experiment (Salaita et al., 2005). Careful evaluation of the exact timing of radicle protrusion between the mutant putatively exhibiting faster completion of germination and the wild type was conducted from the chronology of images.

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