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First published online 10 June 2003
doi: 10.1242/jcs.00615

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Restricted spatial expression of a high-affinity phosphate transporter in potato roots

¹ Rothamsted Research, West Common, Harpenden, Herts, AL5 2JQ, UK
² Research Centre for Eco-environmental Sciences, The Chinese Academy of Sciences, Beijing 10085, China
³ Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476 Golm, Germany

View larger version (26K): [in a new window]   Fig. 1. StPT2 is present in the plasma membrane fraction of potato roots. Western blotting of plasma membrane preparations from P-fed and P depleted potato roots and leaves to show presence of two immunolabelled bands that are reduced in intensity in P-fed plants and absent from leaf extracts. 20 µg of protein was loaded on each lane.

View larger version (30K): [in a new window]   Fig. 2. Expression of StPT2 is predominantly in the root tip and is increased by P depletion. Measurement of StPT2 in tip (1 cm), middle (next 4 cm) and upper (root base) parts of P-depleted and P-supplied roots by (A) western blotting of whole root extracts probed with StPT2 antibody and (B), RQ RTPCR using StPT2 gene specific primers with C, 18 S rRNA using 18S specific primers as a loading control.

View larger version (86K): [in a new window]   Fig. 3. Whole-mount immunolocalisation indicates that StPT2 is present in the elongation zone where it is confined to the epidermal layer, and in the root hairs. (A-E) Superficial staining pattern in root tips, and F shows optical section collected at 100 µm depth. (Arrow indicates root cap cells.) (G) Projected image of quartz-grown plants to show labelling in the root hairs. Roots were excised from P-depleted (A, C-G) or P-supplied (B) plants and labelled with -tubulin (A-F) antibody (green) and StPT2 (A,B,F,G), pre-immune (C) VPPase (D) and ATPase (E) antibodies (red). Yellow indicates colocalisation of red and green. Bar, 100 µm.

View larger version (164K): [in a new window]   Fig. 4. StPT2 is outside -tubulin labelling but inside cell wall labelling. P-depleted root whole-mount tissue from the tip region viewed at a depth of 10 µm, labelled with -tubulin (A-C), cell wall (D) (green) and StPT2 (A and D), plasma membrane ATPase (B) and VPPase (C) (red). In A and B, dotted arrow (CS, cross-section) indicates cell with peripheral red labelling where the optical section cuts across a region of the cell with label around its circumference. Single arrow (EF enface), cell with red labelling covering the green labelling where the optical section passes above the cell. Double arrow indicates cell without StPT2 labelling. In D, solid arrow is plasma membrane (PM) and dotted arrow, cell wall, (CW). Bars, A and B, 25 µm; C and D, 16 µm.

View larger version (62K): [in a new window]   Fig. 5. Image of root transverse sections showing that StPT2 labelling is confined to the apical plasma membrane of epidermal cells, whereas the plasma membrane ATPase labels the whole cell periphery of epidermal and outer cortical cells. -Tubulin labelling is across the whole root. Transverse sections (50 µm) were taken of root whole mount 700 µm from the tip (apart from C, 300 µm) labelled with StPT2 (A,C,D,F) and plasma membrane ATPase (E,G) (red) and -tubulin (green) (B,C,F,G). Double dotted arrow indicates labelling with StPT2 of the apical plasma membrane of the epidermal cells. Single arrowhead shows some ATPase labelling in the cytoplasm; double arrow shows possible enface view of a cell. Bars, A and B, 50 µm; C, 100 µm; D-G, 16 µm.

View larger version (10K): [in a new window]   Fig. 6. Relative fluorescence intensity in basal (black) and apical (white) plasma membranes of epidermal cells 700 µm from the tip labelled with StPT2 and plasma membrane ATPase antibodies detected with Alexa Fluor 568 goat anti-rabbit secondary antibodies. Results show mean±s.e. for 10 replicates.

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