Supplemental Figure S1

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1 Supplementl Figure S1 TG nrt1.5- Li et l., 1 nrt1.5- Lin et l., 8 F L CTGCCT R T 5'UTR 3'UTR p (k) nrt1.5- C nrt1.5- Supplementl Figure S1. Phenotypes of the T-DN insertion mutnts (this pper), nrt1.5- (Li et l., 1) nd nrt1.5- (Lin et l., 8)., NRT1.5 gene structure nd T-DN insertion sites of the mutnt lines nrt1.5-, nrt1.5- nd. Open oxes: untrnslted regions (UTRs). Filled oxes: exons. rrows: positions of PCR primers used for the identifiction of., Representtive picture of, nd nrt1.5- plnts grown on low fertilized soil s shown in Fig. 1. C, Representtive picture of, nd nrt1.5- plnts grown on low fertilized soil s shown in Fig. 1.

2 Supplementl Figure S DS 3 DS 57 DS C D E rosette FW [g] F G H nitrte [mg g FW ] * 3 57 DS chlorophyll + [µg mg FW ] totl N [%] * Fv/Fm DS DS nthocynins [µg mg FW ] totl C [%] DS 3 57 DS 3 57 DS 3 57 DS I J K L Seed yield [mg plnt ] 3 1 * 1 Seed weight [mg] * Oil per seed [%] Protein per seed [%] Supplementl Figure S. Supplementtion with 1 mm KNO 3 suppressed the pleiotrophic phenotype on low fertilized soil., Phenotype of nd rosettes on low fertilized soil supplemented with 1 mm KNO 3. Plnts t, 3 nd 57 dys fter sowing (DS) re shown. In three repets of the experiment plnts developed the sme phenotype., PS II mximum quntum efficiency decline in lef no. 8. lck nd grey rrows: first decline in Fv/Fm in nd plnts, respectively, indicting senescence initition (mens ± SD; n = 8). C, Rosette fresh weight of nd t, 3 nd 57 DS. Dt re mens ± SD (n = 8). D, Chlorophyll concentrtions in nd rosettes. Dt re mens ± SD (n = 3). E, nthocynin ccumultion in nd leves. Dt re mens ± SD (n = 3). F, Nitrte (mens ± SD; n 5), G, Totl N (mens ± SD; n ) nd H, Totl C (mens ± SD; n ) concentrtions in homogenized rosette mteril of nd plnts t, 3 nd 57 DS. I, Quntifiction of seed yield, J, 1 seed weight, K, oil content nd L, protein content in dry seeds of nd plnts grown on low fertilized soil, supplemented with 1 mm KNO 3. Dt re mens ± SD (n = 8). * nd ** indicte significnt differences (Students t-test) etween nd with p <.5 nd p <.1, respectively. Similr results were otined in two independent experiments.

3 Supplementl Figure S3 ction conc. [meq g DW ] 3 1 leves 1 leves 13 roots inflorescence stem Mg C K Supplementl Figure S3. ltered ction distriution in plnts. Concentrtions of the three mjor ctions K +, C + nd Mg + in pooled old leves no. 1, pooled young leves no. 13, roots nd inflorescence stems of hydroponiclly grown nd plnts shown in Fig.. Milliequivlents were clculted sed on the elementl concentrtions shown in Tle 1, respecting the tomic weight nd vlence of the three ctions. Dt re mens ± SD (n 9).

4 Supplementl Figure S Jsmonic cid metolism iosynthesis Signlling Defenserelted Ctolism 1 Reltive expression [log FC] Clcium Miscellneous Signlling Trnsport Defense Secondry Metolism ROS Polymine iosynthesis Lectins Chitinse 16 1 Reltive expression [log FC] Supplementl Figure S. Differentilly regulted iotic stress response nd J signlling genes in leves of plnts., Reltive trnscript levels of genes relted to J metolism nd, 1 genes relted to clcium signlling nd other plnt stress response pthwys in leves (no. 13) of nd wild-type plnts grown in hydroponic culture were mesured y qpcr nd normlized to UQ1 (mens ± SD, n = ). Plotted re the log -fold expression chnges in leves compred with leves.

5 Supplementl Figure S5 6. shoot 6. root C [mg x g DW] scion: stock: 8. shoot 8. root Mg [mg x g DW] c c scion: stock: Supplementl Figure S5. C nd Mg concentrtions in shoots nd roots of grfted ridopsis plnts. Grfted plnts were grown for six weeks on unfertilized 'type ' soil. For the first two weeks the soil ws supplemented with 1 mm KNO 3 to support plnt growth. Finlly, phenotype development ws provoked y fertiliztion with 1 mm KNO 3., C nd, Mg concentrtions in shoots nd roots of grfted plnts s indicted elow the rs (mens ± SD; n 6). The dt were sttisticlly nlyzed y one-wy NOV nd susequent multiple comprisons (Tukey s HSD test). Different letters indicte significnt difference t p <.5. Verticl rs denote stndrd devitions.

6 Supplementl Figure S6 Control nrt1.5- PHO1p::NRT1.5 #1 PHO1p::NRT1.5 #3 1 mg/l Hyg C 5 mm KCl Supplementl Figure S6. Root growth phenotypes of nrt1.5 mutnts nd PHO1p:NRT1.5 complementtion lines in response to Hyg nd high potssium concentrtion. Seedlings pre-germinted for four dys on.5 x MS medium were trnsferred on,.5 x MS medium s control,, medium supplemented with 1 mg/l Hyg, C, medium supplemented with 5 mm KCl, where they continued to grow verticlly for 6 more dys.

7 Supplementl Figure S7 skor-3 TG F R T 5'UTR CHC cnd nky 3'UTR 1 3 / gdn 3,881 k SKOR NRT1.5 GPC x x x C skor-3 N/K/P [mm] 1/1/1 1/1/1 /1/1 Supplementl Figure S7. Identifiction of the T-DN insertion mutnts nd skor-3 in the ckground nd confirmtion of the sence of full-length trnscripts in nd the doule mutnt /., T-DN insertion sites in the SKOR genomic region of the mutnt lines nd skor-3 used in this study. The T-DNs re inserted in the eighth nd second exon of the gene, respectively. CHC, chnnel hydrophoic core; cnd, cyclic nucleotide-inding domin; nky, nkyrin domin. lck nd white oxes represent exons nd untrnslted regions (UTRs), respectively. The rrows indicte the nneling positions of the forwrd (F) nd reverse (R) primers used for PCR on genomic nd cdn., RT-PCR ws performed with gene-specific primers for SKOR on ng nd for NRT1.5 nd GPC on 5 ng reverse-trnscried totl RN from seedlings s indicted on top. PCR cycle count ws. Genomic DN (gdn) ws used s control. C, Phenotypes of,, nd skor-3 under different fertiliztion regimes.

8 Supplementl Figure S8 N/K/P [mm] /1/1 N/K/P [mm] 1/1/1 K [mg x g DW] c / c / C [mg x g DW] Mg [mg x g DW] / / Supplementl Figure S8. Rosette phenotype nd K, C nd Mg concentrtions in,, nd / plnts following /1/1 nd 1/1/1 mm N/K/P supply., K, C nd Mg tissue concentrtions in the rosettes re represented s rs for (light grey), (drk grey), (white) nd / (lck), following the fertiliztion with /1/1 mm N/K/P. The phenotype of the rosettes is shown t the ottom., K, C nd Mg concentrtions s in, ut following the tretment with 1/1/1 mm N/K/P. The phenotype of plnts is shown t the ottom. The dotted red line indictes K concentrtion of 1% in the dry mtter. Sttisticl nlysis of the dt ws performed y onewy NOV nd susequent multiple comprisons (Tukey s HSD test). Mens (n 6) mrked with different letters differed significntly t p <.5. Verticl rs denote stndrd devitions.

9 Supplementl Figure S9 Reltive trnscript level 8 6 N/K/P [mm] 1/1/1 1/1/1 1/1/1 1/1/1 tnrt1.5 tskor Supplementl Figure S9. Expression of NRT1.5 nd SKOR under different fertiliztion regimes. Reltive trnscript levels of NRT1.5 nd SKOR were mesured y qpcr nd normlized to UQ1 in roots of plnts grown on non-fertilized soil supplemented with different fertiliztion solutions (mens ± SD; n ). Reltive trnscript levels of NRT1.5 nd SKOR in roots grown in soil supplemented with 1/1/1 mm N/K/P re set to 1..

10 Supplementl Figure S1 skor Ws Ws nrt1.5- N/K/P [mm] 1/1/1 1/1/1 1/1/1 1/1/1 1/1/1 5//5 Supplementl Figure S1. Phenotypes of ridopsis nd Wssilewskij [Ws] ecotypes nd different nrt1.5 nd skor mutnts under different fertiliztion regimes., Nine plnts ech of the lines skor[ws], Ws wild type, nrt1.5-[], [], wild type nd [] were rrnged on soil s indicted., On low fertilized soil supplemented with the N/K/P nutrient solutions indicted t right nd Ws wild type plnts developed different phenotypes, nd lso the phenotypes of the two skor mutnts in nd Ws ckgrounds differed.