Supplementary information of manuscript Bonke/2003-07-07358A
Supplementary file types:
supplementary S1.jpg (269 kb)
supplementary S2.jpg (218 kb)
supplementary S3.jpg (168 kb)
supplementary S4.jpg (213 kb)
Supplementary Information figure legends
Figure S1a. Cross sections of 9 days old wild- type and apl petioles. Insets showing close-up of the phloem poles. Scale bar 20 mm.
Figure S1b. Cross sections of 13 days old wild-type and apl hypocotyls. Insets showing close-up of the phloem poles. Protoxylem cells are indicated with black arrows. Scale bar 20 mm.
Figure S2a. Early protophloem SE differentiation is not taking place in apl (visualized by toluidine blue staining in root sections 70 mm above the QC). Scale bar, 30 mm. PP – protophloem SE, P – pericycle.
Figure S2b. Phloem initial marker line J0701 crossed with xylem axis initial marker line J1721. Schematic represents the phloem expression pattern of J0701. Compressed series of confocal images. Scale bar, 60 mm.
Figure S2c. J0121: GFP expression in pericycle cells associated with the protoxylem cells in wild-type and apl, respectively. Compressed series of confocal images Scale bar, 60 mm.
Figure S2d. J1721: GFP expression in the protoxylem cells in the root meristem in wild-type and apl, respectively. Compressed series of confocal images Scale bar, 60 mm.
Figure S3a. APLpro::GUS expression is also observed in phloem of true leaves (left). Cross section of the root shows the protophloem SE specific expression pattern as seen with in situ hybridization (right). P; pericycle, X; xylem axis.
Figure S3b. GFP-APL expression as conferred under the APL promoter is first found in the nucleus of the developing protophloem SE at approximately 6-8 cells above the phloem initial cell (left panels and the schematic tracing displaying the same colour code as that in Fig. 1.)
Figure S3c. Cross-section of hypocotyl of a late-torpedo-stage embryo. The same characteristic pattern of four nearly identical poles can be found as in embryonic root later during embryogenesis. This vascular stage precedes patterning of phloem and APL expression during late embryogenesis, after the patterning of the outer tissues is completed.
Figure S4a. WOLpro::GFP expression pattern shows strong vascular expression in the root meristem, which fades out higher up.
Figure S4b. Close up confocal image of the WOLpro::GFP root showing the procambial specific expression. Scale bar, 60 mm .
Figure S4c. In situ hybridization with the APL probe in a WOLpro::APL transgenic line shows procambial specific expression proximal to the root meristem. Scale bar 20 mm (right).
Figure S4d. Cross section approximately 70 mm above the QC shows that the cellular pattern in the WOLpro::APL line is essentially identical to that in wt (compare FigS2a). PP – protophloem SE, P – pericycle.
Figure S4e. In WOLpro::APL line we don’t observe differentiation of TE in protoxylem position and a delayed differentiation of TE in metaxylem position. In this view, the lack of TE in protoxylem position can also be observed from the fact that the characteristic annular cell wall thickenings in the marginal cell files are missing, when TE are finally starting to appear in the metaxylem position in the WOLpro::APL plants. Lack of xylem appears to correlate with the high activity of the APL promoter.
Figure S4f, g. Toluidine blue (f) and DAPI stained (g) images shown as an overlay in Figure 4e.
Figure Sh, i. Toluidine blue (h) and DAPI stained (i) images shown as an overlay in Figure 4f.