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Subcellular Localization of NRT1.8. Onion epidermal cells transiently transformed with either the NRT1.8::EGFP fusion or unfused EGFP were incubated in 0.8 M mannitol to induce plasmolysis and then imaged by confocal microscopy. (A) Fluorescence image of epidermal cell expressing the NRT1.8::EGFP fusion protein. (B) Merged EGFP fluorescence and bright-field image. (C) Fluorescence image of epidermal cell expressing EGFP as a control. (D) Merged control EGFP fluorescence and bright-field image. (E) and (F) Arabidopsis protoplasts expressing the NRT1.8::EGFP protein (E) and bright-field image (F). The yeast strain YNVW1 (Δdur3, Δura3) was unable to grow on 2 mM urea as a sole nitrogen source. YNVW1 was transformed with the yeast expression vector pHXT426 (Wieczorke et al., 1999) alone or harboring the open reading frame of AtDUR3 and plated on yeast nitrogen base without ammonium and amino acids agar (Difco) adjusted to pH 5, 6, or 7. Localization of the promoter activity of AtDUR3. Transgenic plants expressing an AtDUR3-promoter:GFP construct were grown on MGRL agar plates under nitrogen-sufficient (+N) or nitrogen-deficient conditions (?N). Images from root tips (e and f) or lower (c and d) and upper (a and b) root hair zones were taken by an apotome-equipped fluorescence microscope. Bars represent 50?μm. rh, ep, co and xy indicate root hair, epidermis, cortex and xylem, respectively. AtDUR3 localizes to the plasma membrane. (b) Protein gel blot analysis of root membrane fractions from roots of wild-type (Col-0) plants cultured under a continuous supply of 2?mm ammonium nitrate (+N) or under nitrogen deficiency for 1, 2 or 3?days (?N). (d) Intracellular localization of the AtDUR3 protein in nitrogen-starved root hairs from Col-0 (a and c) or from atdur3-1 (b and d) plants (1) morphology modification -------root hair Two accessions of Arabidopsis thaliana, that were very efficient in phosphate acquisition (1) morphology modification -------Lateral root
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