Fig. 4: GmPHF1 interacts with GmPT4 for P uptake in soybean roots.

a FPKM values of PT family genes in soybean roots (3-week-old) revealed by RNA-seq. b Expression levels of GmPT4 in different tissues of soybean revealed by real-time RT-PCR. Data are presented as mean values±SD of three biologically independent samples for each tissue. c V and Rb uptake assay in transgenic hairy roots. p35S:GmPHF1, p35S:GmPT4, and Cas9-GmPT4 constructs were individually introduced into the hairy root of WT (YC04-5) or GmPHF1-RNAi line. The receptor parents of transgenic hairy root are indicated in parentheses. Root samples were treated with V (1 μM, Pi tracer) and Rb (1 μM, internal control) for 12 h. P values were calculated using Kruskal–Wallis test. Different letters indicate a significant difference in element uptake (P < 0.05). Data are presented as mean values ± SD. n = 32, 30, 23, 9, 24, 14, 10 biologically independent samples, respectively. d Growth of wild-type plant (WT), overexpression line of GmPHF1 or GmPT4, co-overexpression line of GmPHF1 and GmPT4 at seedling stage (20-d-old plants). Bar = 50 cm. e, f Phenotypic parameters including plant biomass (dry weight, (e)) and P-acquisition efficiency (P uptake per unit root length, (f)) of the WT and three overexpression lines in d. Data are presented as mean values ± SD. n = 5, 6 biologically independent samples in e, f, respectively. P values are 1.59 × 10⁻⁰² in e and 1.03 × 10⁻⁰² in f based on Kruskal–Wallis test and different letters indicate significant differences (P < 0.05). g Immunostaining of GmPT4 localization in WT and GmPHF1-RNAi line. Immunostaining was performed with ProPT4: gPT4-GFP transgenic hairy roots. Red fluorescence is the signal from the GFP antibody and Cyan fluorescence is the signal from the cell wall. Bars = 20 μm. Data are presented as mean values + SD of ten biological replications. P value was calculated based on two-sided t-test. Source data underlying panels a–c, e–g are provided as a Source Data file.