Figure S1. Si uptake by the whole roots in two pumpkin cultivars. (A) Time-dependent uptake of Si. Seedlings of 14-d-old were exposed to a solution containing 0.5 mM silicic acid for different times. (B) Concentration-dependent uptake of Si. Seedlings were exposed to a solution containing different concentrations of Si . (C) Si concentration in the xylem sap. Seedlings exposed to different Si concentrations for 24 hours were decapitated and the xylem sap was collected for 30 min. All data are means of three biological replicates.

Figure S2. Phylogenetic tree of silicon influx transporters in pumpkin (Cm-), rice (Os-), maize (Zm-), barley (Hv-) and zucchini CpNIP1. The 0.1 scale shows substitution distance.

Figure S3. Alignment of amino acid sequences of silicon influx transporters from rice (Os Lsi1) and two pumpkin cultivars (B+, Shintosa; B, Super-unryu). Green boxes show amino acid residues composing ar/R selectivity filter and gray shadow shows NPA motifs. Pink box shows amino acid residue difference between two pumpkin cultivars

Figure S4. Localization of GFP-CmLsi1 fusion protein expressed in Xenopus oocyte. (a), (d) water injected control. (b), (e) GFP-CmLsi1(B+). (c), (f) GFP-CmLsi1(B+P242L). (a-c) confocal image of intact oocyte. (d-f) squashed oocyte. GFP-CmLsi1(B+) and GFP-CmLsi1(B+)P242L were digested from the vectors which were used for the subcellular localization assay, with XbaI and PstI. Then these fragments were inserted into oocyte expression vector pXßG-ev1. Oocytes injected with cRNA were squashed between the slide grass and cover grass, and GFP fluorescence was observed by laser-scanning confocal microscopy (LSM700; Carl Zeiss).

Appendix S1. Alignment used to generate the phylogenetic tree presented in Figure S2.

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