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Figure S1. Ion concentrations of guttation fluid from the hydathode and xylem exudates along the longitudinal axis of primary barley leaf grown for 9 d under high humidity. (a) Ca2+ and (b) Mg2+ were measured using ion chromatography. The x-axis indicates the relative position along the leaf from 0% at the base to 100% at the tip. Plants were grown in full strength culture medium (closed circles) or in one-tenth culture medium (open circles). Values are expressed as mean ± SE [n = 3–10 (xylem exudates) to 9–15 (guttation fluid: symbols are different from xylem exudates)]. Datasets in which there is a significant change along the leaf (P < 0.05) are indicated with an asterisk.

Figure S2. Fluorescent X-ray imaging of elemental K. PL: primary leaf, SL: second leaf. Barley plants were grown in one-tenth culture medium (a) or in full-strength culture medium (b). Bright parts show higher concentrations of K.

Figure S3. Tissue ion concentrations based on fresh weight basis [(a) Ca2+ and (b) Mg2+] along the longitudinal axis of a primary leaf of barley grown for 9 d under high humidity. The x-axis indicates the relative position of leaf length from 0% at the base to 100% at the tip. Plants were grown in full-strength culture medium (closed circles) or in one-tenth culture medium (open circles). Values are expressed as mean ± SE (n = 3–8).

Figure S4. Autoradiogram of 32P distribution in a barley plant grown for 8 d. Leaves were incubated in a solution containing 32Pi for 15 min: (a) intact leaf and (b) leaf whose lower epidermis was peeled. In each panel, left is an autoradiogram, and right is a photograph of the plant. 32Pi concentration is indicated by pseudo colour from high (white) to low (blue) or absent (black).

Figure S5. Comparison of osmolality at different positions along the primary leaf of barley grown for 8 d under high (H) or low (L) humidity. X-axis indicates the relative position along the leaf from 0% at the base to 100% at the tip. Osmotic values are expressed as mean ± SE (n = 7). The asterisk above the H-100 treatment indicates that it is statistically different (P < 0.05) from the other treatments.

Figure S6. Expression levels of Pi, K+ and NO3 transporter genes along the primary leaf of barley analysed with real-time PCR. (a) Pi transporters: HvPT1, HvPT3, HvPT4, HvPT5, HvPT6, HvPT7 and HvPT8. (b) K+ transporters: HvHAK3 and HvHAK4. (c) NO3 transporters: BCH1 (HvNRT2.1) and BCH2 (HvNRT2.2). The expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene (d) was used as the control and relative expression normalized to the expression in the tip (100%). Measurements were repeated two to three times. Datasets in which there is a significant change along the leaf (P < 0.05) are indicated with an asterisk.

Figure S7. Ion concentration differences between tissues and xylem exudates of NO3 (closed circles), K+ (open circles), Pi (closed triangles) and Cl (open triangles) along the longitudinal axis.

Figure S8. The photosynthetic activity of a detached primary leaf measured using chlorophyll fluorescence imaging. (a) The maximum quantum yield of PSII photochemistry (Fv/Fm) (open circle) and apparent quantum yield (ΦPSII) (closed circle) at each position in a leaf. Values are expressed as mean ± SE (n = 4). (b) An example of chlorophyll fluorescence images of barley primary and second leaves. False colour index is shown at the side.

Table S1. Primers used for real-time RT-PCR.

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