Fig. 4: Mutation of the AtPHR1 KHR motif impairs interaction with AtSPX1 and Pi homeostasis in planta. | Nature Communications

Fig. 4: Mutation of the AtPHR1 KHR motif impairs interaction with AtSPX1 and Pi homeostasis in planta.

From: Inositol pyrophosphates promote the interaction of SPX domains with the coiled-coil motif of PHR transcription factors to regulate plant phosphate homeostasis

Fig. 4

a Growth phenotypes of Col-0 wild type, phr1-3 of phr1-3 complementation lines expressing FLAG-AtPHR1, FLAG-AtPHR1K325A, FLAG-AtPHR1H328A, FLAG-AtPHR1R335A, FLAG-AtPHR1K325A R335A, and FLAG-AtPHR1K325A H328A R335A under the control of the AtPHR1 promoter at 21 DAG grown in Pi-sufficient conditions. One representative line per complementation construct is shown (specified by a #), additional lines are shown in Supplementary Fig. 4. b Plot representing the cellular Pi content of the lines shown in a. Bold black line, median; box, interquartile range (IQR); whiskers, lowest/highest data point within 1.5 IQR of the lower/upper quartile. Seedlings were germinated and grown on vertical 1/2MS plates for 8 days, transferred to 1/2MS plates supplemented with either 0, 1, or 10 mM Pi and grown for additional 7 days. For each line, four plants were measured in technical duplicates. Pi contents of all lines can be found in Supplementary Fig. 4 (*p < 0.05; **p < 0.01; ***p < 0.001). c Heat maps of PSI marker gene (ACP5, IPS1, MGD3, PECP1, PPsPase, SPX1) expression analyses of the lines shown in a, represented as Z-scores. For each line, three biological replicates were analyzed in technical triplicates by qRT-PCR. d Co-immunoprecipitation (Co-IP) experiment assessing the ability for immobilized GFP-AtPHR1 and GFP-AtPHR1KHR/A to interact with mCherry-AtSPX1 in N. benthamiana. Input western blottings are shown alongside. Experiment was performed twice with similar results. e Genetic interactions in the VIH-PHR signaling pathway. Col-0 wild type and the indicated mutant seedlings were grown on 1/2MS plates for 7 DAG, transferred to 1/2MS plates supplemented with either 0, 1, or 10 mM Pi and grown for additional 7 days. For complementation analyses, wild-type AtPHR1 or AtPHR1KHR/A was expressed as an N-terminal eGFP fusion protein under the control of the AtPHR1 promoter. f Heat maps of PSI marker gene expression for the lines shown in e.

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