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Supplemental Data. Møller et al. (2009) Shoot Na + exclusion and increased salinity tolerance engineered by cell type-specific alteration of Na + transport in Arabidopsis Supplemental Figure 1.

1 Supplemental Data. Møller et al. (2009) Shoot Na + exclusion and increased salinity tolerance engineered by cell type-specific alteration of Na + transport in Arabidopsis Supplemental Figure 1. Salt-sensitive phenotype of salt-stressed Pro35S:AtHKT1;1 plants. Pro35S:AtHKT1;1 plants (center and bottom) are more salt-sensitive than the Col-0 wild-type plants (top) and show stunted growth and chlorosis. Plants were grown as outlined for Figure 5 with a final NaCl concentration of 50 mm.

2 J2731* LB GFP UAS GAL4 RB At1g bp 1088 bp At1g21050 E2586 LB GFP UAS GAL4 RB At1g bp 3964 bp At1g70760 Supplemental Figure 2. Position of enhancer trap T-DNA in the genome of J2731* and E2586. Diagrammatic representation of the site and orientation of insertion of the T- DNA inserts in the enhancer trap lines, as determined by TAIL-PCR. The adjacent genes are annotated as follows. At1g21040 is predicted to be a pseudogene with similarity to genes encoding mutator-like transposases. At1g21050 is expressed but has unknown function. At1g70750 is expressed but has unknown function. At1g70760 is expressed and has similarity with an inorganic carbon transport protein from cyanobacteria.

Photographs illustrating the laser dissection of a

to separate cells in the stele and cells of the outer

(A) Fixed transverse section of a root before

(B) Endodermal cells halfway through being removed by

(C, D) The disc of cells comprising the stele was cut

3 A B C D E F G H Supplemental Figure 3. Laser dissection of transverse root sections. Photographs illustrating the laser dissection of a transverse section of an J2731* UASGAL4:AtHKT1;1 root to separate cells in the stele and cells of the outer part of the root. (A) Fixed transverse section of a root before commencement of laser dissection. (B) Endodermal cells halfway through being removed by the laser. (C, D) The disc of cells comprising the stele was cut out and has fallen into a PCR tube below the microscope stage. (E, F) The remaining outer part of the root consisting of the epidermal and cortical cells was cut out with the laser. (G, H) The outer part has been cut out and has fallen into a PCR tube below the microscope stage. For A-C, E-G scale bars indicate 50 µm.

4 Ph Co En Pe Xy Supplemental Figure 4. Electron micrograph of transverse section of J2731*UASGAL4:AtHKT1;1 root. The anatomy of root cells of J2731* and J2731*UASGAL4:AtHKT1;1 plants was examined with transmission electron microscopy. Plants were grown under the same conditions as for x-ray microanalysis. The tissue analysed was located 20 mm from the root tip. No structural differences in terms of cell size, number and shape or degree of vacuolation were seen between genotypes (data not shown). Co, cortex; En, endodermis; Pe, pericycle; Xy, xylem; Ph, phloem. Scale bar indicates 10 µm.

5 Genotype B Na Mg P S K Ca Mn Fe Zn J2731* parents n = 93 J2731* nulls n = 71 J2731* UAS:HKT N = 285 E2586 parents n = 65 E2586 nulls n = 39 E2586 UAS:HKT N = ± ± ± ± ± ± ± ± ± ± ± ± 22 a 376 ± ± 19 a 631 ± ± 64 a 1052 ± ± ± ± ± ± 6 b 374 ± ± 8 b 590 ± ± 3 b 1090 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 21 a 429 ± ± ± 15 a 4819 ± 96 a 1173 ± ± ± ± ± ± 4 b 372 ± ± ± 6 b 5143 ± 38 b 1101 ± ± ± ± 0.1 Supplemental Table 1. Shoot accumulation of ten elements in UAS GAL4 :HKT1;1 plants. Using ICP-AES, the content in mg. kg -1 FM of ten elements was measured in leaf tissue from five-weekold plants grown on a soil-like mix and supplemented with a nutrient solution containing 2 mm NaCl. Segregating J2731* UAS GAL4 :HKT1;1 T2 plants descending from 19 independent T1 plants and segregating E2586 UAS GAL4 :HKT1;1 T2 plants descending from 13 independent T1 plants were used in this experiment. All UAS GAL4 :HKT1;1 plants were genotyped using PCR to detect the presence or absence of the T-DNA insert. Statistical analysis was applied for the comparison of the transgenic UAS GAL4 :HKT1;1 plants with the corresponding null segregants. The data were log transformed and an F-test was performed with P < 0.05 to determine equal or unequal variance in the data for each element. Accordingly, a two-sample two-tailed Student s t-test was performed (P < 0.01) assuming equal or unequal variance. Values that were found to be significantly different using the described Student s t-test with P < 0.01 are indicated with a and b. When comparing the null segregants with the corresponding parental enhancer trap line, no elements were found to be significantly different using the same statistical analysis. Values presented are averages ± SEM.

6 Genotype Shoot [Na + ] Root [Na + ] Shoot [K + ] Root [K + ] J2731* 5133 ± ± ± ± 120 J2731* UAS:HKT 2600 ± ± ± ± 133 Supplemental Table 2. Accumulation of Na + and K + in shoot and roots of two-week-old J2731*UAS GAL4 :HKT1;1 plants. The content of Na + and K + in whole shoot and whole root tissue of two-week-old seedlings grown in hydroponics as for x-ray microanalysis was measured using ICP-AES. The tissues were separated, rinsed in MilliQ water, blotted and weighed prior to ICP-AES analysis. Three replicates, each with nine shoots or root systems, were used. Values presented are averages ± SEM.

Supplemental Figure 1: Increased Fe deficiency gene expression in roots of nas4x-2

Supplemental Figure 1: Increased Fe deficiency gene expression in roots of nas4x-2 IRT1, FRO2 and FIT expression levels in roots of the wild-type, nas4x- 1 and nas4x-2, showing that in both nas mutants