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Extra info for Environmental Chemistry: New Techniques and Data
All hydroponic solutions were continuously aerated by an electric pump. At T0 stage and after a period of 6, 30, and 54 hr plants were harvested, washed with distilled water and then blotted with paper towels. Finally, roots and leaves were separated and the samples were frozen in liquid N2 and stored at –80°C. 4 mM CaSO4 solution for 15 min for removing the anion present in the apoplast before sampling. Levels of Nitrate Nitrate was extracted from the tissues by homogenizing the samples previously boiled in 4 volumes of distilled water for 15 min.
The homogenate was centrifuged at 13,000 g for 20 min at 4°C to obtain a clarified supernatant. Chlorophyll concentration was measured according to Lichtenthaler . Determination of Malondialdehyde and Chlorophyll Fluorescence of the Leaves Malondialdehyde (MDA) was assayed by the method of Heath and Packer . 1% (w/v) trichloroacetic acid (TCA) and centrifuged at 13,000 g for 20 min at 4°C. 5% (w/v) thiobarbituric acid was added to the supernatants, which were subsequently heated at 95°C for 30 min.
05), coupled with a threshold of 2-fold change in the amount, revealed that 20 spots in roots and 18 spots in leaves were affected by nitrogen availability. Figure 4. The 2-DE maps. Representative 2-DE maps of soluble protein fractions extracted from roots (A) and leaves (B) of Zea mays plants. 5% SDS-PAGE and visualized by Coomassie Brilliant Blue G-250 (cCBB)-staining. 05) after the exposure to 10 mM nitrate for 30 hr. The analysis of these spots by LC-ESI-MS/MS allowed to identify 15 and 14 proteins in root and leaf patterns, respectively.