Fig. 1. Analysis of the sulphate transporter gene family 3 (SULTR3) in rice. A, Expression of OsSULTR3 family genes in wild type (WT, Nipponbare) and Osphr1Osphr2Osphr3 triple mutants under inorganic phosphate deficiency (-Pi) treatment for 1 d (-Pi1d), 5 d (-Pi5d), 7 d (-Pi7d), 15 d (-Pi15d), and 20 d (-Pi20d). Data are mean ± SD (n = 3). * and ** represent significant differences at the 0.05 and 0.01 levels, respectively, by the Student’s t-test, compared with -Pi0d. B, Subcellular localization of p35S::OsSULTR3.1, p35S::OsSULTR3.2, p35S::OsSULTR3.3, p35S::OsSULTR3.4, p35S::OsSULTR3.5, and p35S::OsSULTR3.6 in rice protoplasts. p35S::OsSULTR3-GFP together with an endoplasmic reticulum (ER)-marker were transiently introduced into rice protoplasts. The green signals indicate GFP, and the red signals indicate ER-marker. GFP, Green fluorescent protein. Scale bars, 10 μm. C, Complementation of the yeast mutant YP100 (Δpho84Δpho87Δpho89Δpho90Δpho91Δgit1) expressing OsSULTR3.2. Equal volumes of 10-fold serial dilutions were applied to YNB (yeast nitrogen base) medium (pH 5.5) with different Pi concentrations and inoculated at 30 ºC for 4 d. PHO84 is a high-affinity phosphate transporter and PHO91 is a low-affinity phosphate transporter, both serves as a positive control, and empty vector as a negative control. D, GUS (β-glucurouidase) staining of the primary root (a), lateral roots (b), lateral root primordium (c), cross section of the GUS-stained primary root (d), GUS-stained leaf blade (e), tissue specific expression of OsSULTR3.2 in whole nodes (f), and enlarged vascular bundles (g). Scale bars, 100 μm for a-c and e-g, and 50 μm for d. E-K, Phenotype (E), biomass (F) and Pi content (G) in shoots and roots, Pi content in different leaves (H), sulfate (S) content in shoots and roots (I), S content in different leaves (J), and Pi content in xylem sap (K) of wild type rice (SSBM) and ossultr3.2 mutants (ossultr3.2-6 and ossultr3.2-10). Seedlings of both the SSBM and ossultr3.2 mutants were excised at the root-shoot junction for xylem sap collection. Leaves 1-5 in H and J refer to consecutive leaves arranged from oldest to youngest in sequence. Data are mean ± SD (n = 3 for F-J; n = 6 for K). In F-K, * and ** represent significant differences at the 0.05 and 0.01 levels, respectively, by the Student’s t-test, compared with SSBM.
Fig. 1. Analysis of the sulphate transporter gene family 3 (SULTR3) in rice. A, Expression of OsSULTR3 family genes in wild type (WT, Nipponbare) and Osphr1Osphr2Osphr3 triple mutants under inorganic phosphate deficiency (-Pi) treatment for 1 d (-Pi1d), 5 d (-Pi5d), 7 d (-Pi7d), 15 d (-Pi15d), and 20 d (-Pi20d). Data are mean ± SD (n = 3). * and ** represent significant differences at the 0.05 and 0.01 levels, respectively, by the Student’s t-test, compared with -Pi0d. B, Subcellular localization of p35S::OsSULTR3.1, p35S::OsSULTR3.2, p35S::OsSULTR3.3, p35S::OsSULTR3.4, p35S::OsSULTR3.5, and p35S::OsSULTR3.6 in rice protoplasts. p35S::OsSULTR3-GFP together with an endoplasmic reticulum (ER)-marker were transiently introduced into rice protoplasts. The green signals indicate GFP, and the red signals indicate ER-marker. GFP, Green fluorescent protein. Scale bars, 10 μm. C, Complementation of the yeast mutant YP100 (Δpho84Δpho87Δpho89Δpho90Δpho91Δgit1) expressing OsSULTR3.2. Equal volumes of 10-fold serial dilutions were applied to YNB (yeast nitrogen base) medium (pH 5.5) with different Pi concentrations and inoculated at 30 ºC for 4 d. PHO84 is a high-affinity phosphate transporter and PHO91 is a low-affinity phosphate transporter, both serves as a positive control, and empty vector as a negative control. D, GUS (β-glucurouidase) staining of the primary root (a), lateral roots (b), lateral root primordium (c), cross section of the GUS-stained primary root (d), GUS-stained leaf blade (e), tissue specific expression of OsSULTR3.2 in whole nodes (f), and enlarged vascular bundles (g). Scale bars, 100 μm for a-c and e-g, and 50 μm for d. E-K, Phenotype (E), biomass (F) and Pi content (G) in shoots and roots, Pi content in different leaves (H), sulfate (S) content in shoots and roots (I), S content in different leaves (J), and Pi content in xylem sap (K) of wild type rice (SSBM) and ossultr3.2 mutants (ossultr3.2-6 and ossultr3.2-10). Seedlings of both the SSBM and ossultr3.2 mutants were excised at the root-shoot junction for xylem sap collection. Leaves 1-5 in H and J refer to consecutive leaves arranged from oldest to youngest in sequence. Data are mean ± SD (n = 3 for F-J; n = 6 for K). In F-K, * and ** represent significant differences at the 0.05 and 0.01 levels, respectively, by the Student’s t-test, compared with SSBM.
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