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For germination assays scoring ABA sensitivity, 20 to 100 seeds per treatment were surface sterillized in 5% hypochlorite and 0.02% Trition X-100. Seed were then rinsed three or four times with sterile water before plating on minmal medium (Haughn and Somerville, 1986), containing 0.7% agar and ABA (mixed isomers; Sigma) at 3 or 5 uM in 15 X 100mm Petri dishes. For scoring kanamycin resistance, the medium included 0.5 X Murashige and Skoog salts (Murashige and Skoog, 1962), 1% sucrose, 0.05% Mes, and 50 ug/mL kanamycin. The dishes were incubated for 1 to 3 days at 4 degrees Celsiu to break any residual dormancy, and then they were tansferred to 22 degrees in continuous light. References: Finkelstein R R, Lynch T J. The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor.[J]. Plant Cell, 2000, 12(4):599-609. |
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