Supplementary Material

10.1071/FP18046_AC CSIRO 2018 Supplementry Mteril: Funtionl Plnt Biology, 2018, 45(11), 1096 1109. Supplementry Mteril Hordeum vulgre nd Hordeum mritimum respond to extended slinity stress displying different temporl umultion pttern of metolites Selm Ferhihi A, Kmel Hessini A,B, Emili Dell Aversn C, Luis D Ameli C, Psqulin Woodrow C, Loredn F. Cirmiello C, Amodio Fuggi C nd Petroni Crillo C,D A Lortory of Extremophile Plnts, Center of Biotehnology of Borj Cedri, University of Elmnr, B.P. 901, Hmmm-Lif 2050, Tunisi. B Biology Deprtment, Fulty of Siene, Tif University, 21974 Tif, P.O. Box 888, Sudi Ari. C Deprtment of Environmentl, Biologil nd Phrmeutil Sienes nd Tehnologies, University of Cmpni Luigi Vnvitelli, Vi Vivldi 43, 81100 Csert, Itly. D Corresponding uthor. Emil: petroni.rillo@unimpni.it

Tle S1. Min physiologil prmeters, inorgni ions, hydrogen peroxide (H2O2), mlondildehyde (MDA), pigments, rohydrtes, glyine etine, proteins, totl nd free mino ids ontent in leves of H. mritimum nd H. vulgre under ontrol (nonsline ondition), slt stress (200 mm NCl) nd slt removl tretment Slt ws grdully dded to slt treted plnts strting from 15 dys fter sowing (DAS). Slt removl tretment strted from 30 DAS. Hrvests were done t 30, 33, 36, 42 nd 48 DAS. Vlues re men s.d. (n = 3)

Hordeum mritimum Hordeum vulgre Dy Control Sl stress Slt removl Control Slt stress Slt removl Fresh weight (g per plnt) 30 1.31 ± 0.07 1.24 ± 0.21 1.24 ± 0.21 2.72 ± 0.31 1.04 ± 0.16 1.04 ± 0.16 33 1.86 ± 0.36 1.25 ± 0.06 1.36 ± 0.06 3.38 ± 0.47 0.87 ± 0.09 d 1.30 ± 0.14 36 2.11 ± 0.11 0.91 ± 0.08 2.08 ± 0.21 4.26 ± 0.39 0.85 ± 0.12 2.22 ± 0.28 42 4.23 ± 0.28 1.24 ± 0.12 2.67 ± 0.48 7.21 ± 0.60 d 0.86 ± 0.11 e 2.72 ± 0.39 48 7.17 ± 0.56 1.41 ± 0.31 4.71 ± 0.44 10.09 ± 0.71 0.86 ± 0.04 d 3.45 ± 0.45 e Reltive wter ontent (%) 30 85.8 ± 7.5 83.8 ± 0.6 83.8 ± 0.6 91.5 ± 3.2 87.0 ± 5.8 87.0 ± 5.8 33 87.0 ± 5.4 83.7 ± 2.2 85.2 ± 4.2 89.7 ± 2.4 86.8 ± 1.6 85.7 ± 3.6 36 87.2 ± 6.8 83.9 ± 3.2 85.9 ± 2.5 89.7 ± 4.0 85.7 ± 2.8 83.0 ± 2.9 42 89.7 ± 2.6 88.6 ± 2.6 89.8 ± 2.5 89.4 ± 2.4 83.8 ± 2.8 90.8 ± 3.6 48 87.3 ± 7.2 87.8 ± 9.0 87.8 ± 1.4 89.6 ± 3.1 83.0 ± 1.5 91.8 ± 1.8 Lef wter potentil (Mp) 30-1.88 ± 0.03-2.47 ± 0.15-2.47 ± 0.15-1.55 ± 0.05-2.27 ± 0.14-2.27 ± 0.14 33-1.80 ± 0.26-2.20 ± 0.17-1.63 ± 0.06-1.52 ± 0.08 d -1.83 ± 0.12-1.40 ± 0.10 d 36-1.53 ± 0.06-2.33 ± 0.06-1.53 ± 0.06-1.30 ± 0.10-2.40 ± 0.10-1.20 ± 0.10 42-1.60 ± 0.10-2.53 ± 0.12-1.57 ± 0.06-1.20 ± 0.01-2.32 ± 0.06 d -1.23 ± 0.06 48-1.50 ± 0.10-2.60 ± 0.17-1.60 ± 0.10-1.17 ± 0.06-2.27 ± 0.06 d -1.20 ± 0.10 Chloride (μmol g 1 FW) 30 63.7 ± 2.5 161 ± 18.6 161 ± 18.6 67.1 ± 1.1 252 ± 9.7 252 ± 9.7 33 65.4 ± 1.1 177 ± 29.5 113 ± 4.5 78.4 ± 4.6 d 277 ± 4.0 e 205 ± 2.0 36 61.7 ± 0.8 206 ± 18.4 93.5 ± 6.1 70.3 ± 4.2 d 274 ± 3.8 e 195 ± 13.6 42 65.0 ± 2.3 206 ± 8.8 65.7 ± 11.5 70.4 ± 3.1 276 ± 6.9 d 127 ± 7.1 e 48 75.6 ± 2.4 197 ± 42.8 61.5 ± 8.8 81.1 ± 16.1 291 ± 17.8 d 109 ± 3.5 e Nitrte (μmol g 1 FW) 30 98.4 ± 5.2 39.2 ± 0.7 39.2 ± 0.7 71.6 ± 4.5 22.4 ± 1.7 d 22.4 ± 1.7 d 33 101 ± 7 37.1 ± 0.7 33.7 ± 4.4 79.0 ± 5.1 19.0 ± 1.3 d 21.8 ± 4.9 d 36 99.1 ± 3.2 42.9 ± 4.4 42.7 ± 8.7 73.3 ± 6.5 17.4 ± 1.1 d 28.8 ± 2.3 e 42 104 ± 8 36.3 ± 0.3 58.8 ± 4.6 73.3 ± 5.3 d 17.3 ± 1.1 e 62.3 ± 3.5 48 99.6 ± 5.8 25.3 ± 1.8 67.6 ± 2.0 71.7 ± 10.3 e 15.9 ± 0.7 d 74.3 ± 2.8 e Potssium (μmol g 1 FW) 30 62.9 ± 1.8 56.2 ± 5.4 56.2 ± 5.4 67.1 ± 1.5 55.0 ± 1.4 55.0 ± 1.4 33 66.7 ± 1.1 54.9 ± 7.1 45.9 ± 3.3 60.1 ± 2.7 50.3 ± 1.9 51.9 ± 0.9 36 63.1 ± 4.6 50.2 ± 5.0 51.7 ± 4.6 63.5 ± 3.7 45.5 ± 2.8 55.2 ± 0.5 42 62.9 ± 4.1 40.7 ± 1.3 56.4 ± 3.9 62.8 ± 1.7 34.3 ± 3.5 d 57.5 ± 0.6 48 57.8 ± 7.4 33.8 ± 3.8 61.5 ± 1.0 63.7 ± 3.2 33.7 ± 1.3 58.6 ± 2.8 Sodium (μmol g 1 FW) 30 19.4 ± 2.6 64.1 ± 7.9 64.1 ± 7.9 12.1 ± 3.9 133.0 ± 9.4 d 133.0 ± 9.4 d 33 20.3 ± 4.9 68.0 ± 2.9 56.3 ± 4.3 13.8 ± 2.2 142.1 ± 5.4 d 125.8 ± 9.4 e 36 23.2 ± 3.7 68.3 ± 4.7 47.3 ± 5.9 13.7 ± 1.7 d 172.7 ± 15.1 e 108.4 ± 6.7 f 42 20.8 ± 4.8 77.5 ± 3.5 31.1 ± 4.4 14.4 ± 2.3 177.5 ± 16.2 d 84.7 ± 4.9 48 24.7 ± 3.1 76.4 ± 2.9 24.6 ± 2.5 15.6 ± 1.2 247.4 ± 26.3 d 65.5 ± 8.3 Potssium:Sodium 30 3.23 ± 0.39 0.88 ± 0.10 0.88 ± 0.11 5.56 ± 0.70 0.41 ± 0.05 d 0.41 ± 0.05 d 33 3.28 ± 0.40 0.81 ± 0.10 0.82 ± 0.10 4.35 ± 0.55 0.35 ± 0.05 d 0.41 ± 0.05 d 36 2.72 ± 0.22 0.74 ± 0.09 1.09 ± 0.11 4.62 ± 0.61 d 0.26 ± 0.03 e 0.51 ± 0.06 f 42 3.02 ± 0.40 0.52 ± 0.05 1.81 ± 0.19 4.36 ± 0.46 d 0.19 ± 0.03 e 0.68 ± 0.08 f 48 2.34 ± 0.33 0.44 ± 0.04 2.50 ± 0.32 4.09 ± 0.50 0.14 ± 0.02 d 0.89 ± 0.10 e Sp osmollity ((mosmol kg 1 ) 30 612 ± 7 823 ± 70 823 ± 70 551 ± 35 1170 ± 235 d 1170 ± 235 d 33 616 ± 4 791 ± 60 644 ± 31 545 ± 58 1045 ± 22 d 1077 ± 212 d 36 530 ± 64 907 ± 80 604 ± 66 533 ± 14 1041 ± 29 766 ± 290 42 640 ± 20 926 ± 27 633 ± 3 523 ± 44 1242 ± 58 d 553 ± 49 48 513 ± 26 1044 ± 44 578 ± 70 471 ± 27 1384 ± 45 d 524 ± 3 H 2 O 2 (μmol g 1 FW) 30 1.16 ± 0.23 1.44 ± 0.22 1.44 ± 0.22 0.75 ± 0.06 2.01 ± 0.37 2.01 ± 0.37 33 0.82 ± 0.26 1.66 ± 0.29 1.53 ± 0.20 0.96 ± 0.12 1.78 ± 0.16 1.25 ± 0.34 36 0.72 ± 0.11 4.65 ± 1.58 1.55 ± 0.26 0.84 ± 0.08 1.54 ± 0.21 1.19 ± 0.11 42 0.69 ± 0.11 5.12 ± 0.95 1.02 ± 0.32 1.11 ± 0.21 1.74 ± 0.58 1.24 ± 0.17 48 0.56 ± 0.19 2.10 ± 0.27 0.76 ± 0.14 0.95 ± 0.06 1.34 ± 0.45 1.06 ± 0.22 MDA (nmol g 1 FW) 30 42.6 ± 3.6 51.8 ± 2.5 51.8 ± 2.5 41.8 ± 7.2 45.7 ± 6.1 45.7 ± 6.1 33 43.1 ± 2.4 50.0 ± 3.2 49.3 ± 2.6 37.2 ± 1.2 44.5 ± 2.5 33.5 ± 4.4 36 43.3 ± 4.5 55.0 ± 3.9 53.6 ± 4.3 d 27.0 ± 5.5 43.5 ± 7.0 d 38.6 ± 4.4 42 42.4 ± 9.8 53.5 ± 6.2 52.0 ± 5.1 31.0 ± 2.3 47.4 ± 4.4 34.3 ± 1.3 48 54.1 ± 5.6 75.8 ± 16.8 57.2 ± 8.3 29.4 ± 6.4 46.1 ± 8.0 35.2 ± 2.1

Hordeum mritimum Hordeum vulgre Dy Control Sl stress Slt removl Control Slt stress Slt removl Totl hlorophyll (mg g -1 FW) 30 2.43 ± 0.01 3.40 ± 0.19 3.40 ± 0.19 2.49 ± 0.25 2.44 ± 0.21 2.44 ± 0.21 33 2.32 ± 0.09 3.14 ± 0.06 3.58 ± 0.25 2.75 ± 0.34 2.30 ± 0.20 1.87 ± 0.26 36 1.91 ± 0.33 d 4.05 ± 0.14 2.88 ± 0.19 2.03 ± 0.57 d 2.22 ± 0.07 1.52 ± 0.32 d 42 1.88 ± 0.09 2.84 ± 0.22 1.59 ± 0.14 2.64 ± 0.16 2.47 ± 0.16 2.42 ± 0.20 48 1.35 ± 0.25 2.42 ± 0.26 1.08 ± 0.01 2.70 ± 0.38 1.38 ± 0.09 1.95 ± 0.21 Chlorophylle /Chlorophylle 30 3.98 ± 0.26 3.68 ± 0.18 3.68 ± 0.18 2.99 ± 0.04 3.19 ± 0.20 3.19 ± 0.20 33 3.55 ± 0.02 3.67 ± 0.13 3.66 ± 0.23 3.12 ± 0.17 3.57 ± 0.25 3.07 ± 0.24 36 4.15 ± 0.65 3.54 ± 0.00 3.70 ± 0.08 3.30 ± 0.02 3.68 ± 0.28 3.30 ± 0.20 42 4.33 ± 0.25 4.01 ± 0.18 4.37 ± 0.39 3.30 ± 0.09 3.41 ± 0.04 3.67 ± 0.18 48 4.14 ± 0.45 4.02 ± 0.16 4.72 ± 0.20 3.69 ± 0.24 3.89 ± 0.19 3.74 ± 0.27 Crotenoids (mg g -1 FW) 30 0.15 ± 0.00 0.20 ± 0.01 0.20 ± 0.01 0.14 ± 0.01 0.14 ± 0.01 0.14 ± 0.01 33 0.14 ± 0.01 0.19 ± 0.01 0.21 ± 0.01 0.16 ± 0.02 0.14 ± 0.02 0.11 ± 0.02 36 0.12 ± 0.02 d 0.24 ± 0.01 0.17 ± 0.01 0.12 ± 0.03 0.13 ± 0.00 0.09 ± 0.02 d 42 0.12 ± 0.00 0.17 ± 0.02 0.10 ± 0.01 0.15 ± 0.01 0.14 ± 0.01 0.14 ± 0.01 48 0.08 ± 0.01 0.15 ± 0.02 0.07 ± 0.00 0.16 ± 0.02 0.08 ± 0.00 0.12 ± 0.01 Strh (µmol gluose eq g -1 FW) 30 24.1 ± 5.1 23.8 ± 2.1 23.8 ± 2.1 10.07 ± 1.51 10.2 ± 1.4 10.2 ± 1.4 33 15.0 ± 0.4 16.2 ± 1.0 17.0 ± 1.4 12.74 ± 1.21 12.0 ± 3.2 11.3 ± 2.8 36 15.3 ± 1.8 13.5 ± 5.1 14.8 ± 1.4 9.92 ± 1.47 13.7 ± 2.8 11.7 ± 1.5 42 11.9 ± 0.5 15.6 ± 1.1 10.8 ± 2.3 10.55 ± 1.52 13.8 ± 3.3 14.7 ± 2.6 48 8.6 ± 1.2 8.2 ± 2.2 9.0 ± 1.9 15.12 ± 3.36 12.7 ± 3.0 15.8 ± 1.3 Gluose (µmol g -1 FW) 30 7.71 ± 0.95 11.18 ± 0.97 11.18 ± 0.97 7.03 ± 1.28 5.05 ± 0.17 5.05 ± 0.17 33 10.67 ± 0.82 8.54 ± 0.87 7.81 ± 0.68 8.99 ± 1.61 6.96 ± 1.38 8.92 ± 1.83 36 6.76 ± 1.20 8.63 ± 0.73 8.40 ± 0.73 9.59 ± 0.94 8.73 ± 0.61 8.06 ± 1.36 42 9.56 ± 0.63 9.44 ± 0.38 6.63 ± 0.78 13.45 ± 4.48 d 12.67 ± 0.21 18.84 ± 3.82 d 48 9.33 ± 1.16 9.63 ± 2.96 8.05 ± 0.82 17.86 ± 1.93 11.69 ± 2.83 13.74 ± 3.34 Frutose (µmol g -1 FW) 30 5.43 ± 0.77 6.68 ± 0.62 6.68 ± 0.62 4.21 ± 0.34 3.48 ± 0.12 3.48 ± 0.12 33 1.48 ± 0.12 1.22 ± 0.19 1.27 ± 0.36 5.16 ± 0.63 4.39 ± 0.53 5.15 ± 0.52 36 0.77 ± 0.14 1.61 ± 0.50 0.96 ± 0.43 5.32 ± 0.21 5.44 ± 0.18 4.43 ± 0.62 42 0.88 ± 0.19 1.54 ± 0.49 0.72 ± 0.11 6.54 ± 1.76 6.60 ± 0.04 9.80 ± 1.39 d 48 1.51 ± 0.28 1.10 ± 0.37 0.76 ± 0.22 7.56 ± 0.50 7.18 ± 1.68 7.17 ± 0.82 Surose (µmol gluoseeq g -1 FW) 7.22 30 3.13 ± 0.75 6.80 ± 1.16 6.80 ± 1.16 3.00 ± 0.21 7.63 ± 0.64 7.63 ± 0.64 33 3.62 ± 0.12 4.95 ± 0.68 d 4.38 ± 0.35 3.94 ± 0.48 7.56 ± 3.54 5.52 ± 0.68 d 36 2.74 ± 0.47 7.02 ± 0.80 4.06 ± 0.87 3.67 ± 0.22 6.72 ± 0.30 3.52 ± 0.12 42 4.34 ± 0.60 7.98 ± 0.35 3.29 ± 0.03 4.23 ± 0.95 14.90 ± 1.69 d 5.92 ± 1.42 48 2.32 ± 0.79 5.48 ± 1.20 1.95 ± 0.72 5.80 ± 0.50 7.18 ± 2.15 4.87 ± 0.31 Glyine etine (µmol g -1 FW) 30 4.99 ± 1.86 12.88 ± 4.42 12.88 ± 4.42 9.04 ± 0.01 15.2 ± 1.2 15.2 ± 1.2 33 6.79 ± 0.21 11.11 ± 0.01 11.42 ± 1.69 5.45 ± 1.40 12.3 ± 0.8 12.1 ± 0.6 36 7.75 ± 1.96 12.74 ± 3.05 8.16 ± 0.33 3.93 ± 0.31 11.9 ± 1.6 10.3 ± 1.7 42 9.32 ± 2.65 11.57 ± 1.61 6.06 ± 1.54 6.18 ± 0.49 11.5 ± 0.9 7.92 ± 2.24 48 5.36 ± 0.59 10.64 ± 1.07 3.64 ± 2.28 6.03 ± 1.36 10.8 ± 2.7 5.23 ± 0.51 Totl proteins (mg g -1 FW) 30 12.0 ± 2.7 12.5 ± 1.5 12.5 ± 1.5 10.4 ± 0.9 9.7 ± 2.7 9.7 ± 2.7 33 10.0 ± 2.9 12.8 ± 4.0 13.4 ± 0.5 9.6 ± 1.4 9.0 ± 3.6 8.2 ± 2.4 36 9.6 ± 3.1 12.7 ± 1.4 13.9 ± 1.2 8.5 ± 0.7 10.1 ± 0.6 8.3 ± 2.7 42 11.2 ± 1.9 14.5 ± 3.6 13.2 ± 3.5 8.5 ± 1.0 10.3 ± 1.6 9.4 ± 2.0 48 11.4 ± 4.0 11.8 ± 3.3 12.8 ± 2.9 8.5 ± 1.4 10.4 ± 1.2 10.7 ± 3.6 Totl free mino ids (µmol g -1 FW) 30 28.6 ± 6.8 33.8 ± 5.9 33.8 ± 5.9 11.5 ± 2.5 54.1 ± 9.5 54.1 ± 9.5 33 25.9 ± 1.2 31.4 ± 5.9 33.7 ± 7.7 12.5 ± 0.2 65.0 ± 7.0 28.7 ± 3.7 36 24.7 ± 6.2 35.1 ± 5.3 26.4 ± 4.7 13.0 ± 4.6 66.9 ± 8.6 24.8 ± 5.0 42 26.4 ± 2.8 47.5 ± 4.7 35.9 ± 3.1 16.1 ± 3.4 d 56.1 ± 8.8 12.7 ± 3.8 d 48 39.6 ± 0.9 36.3 ± 5.7 45.8 ± 3.9 20.2 ± 4.5 48.8 ± 8.0 22.3 ± 1.6

Hordeum mritimum Hordeum vulgre Dy Control Sl stress Slt removl Control Slt stress Slt removl Alnine (mino ids re expressed s µmol g -1 FW) 30 1.39 ± 0.10 1.01 ± 0.29 1.01 ± 0.29 0.70 ± 0.04 1.10 ± 0.08 1.10 ± 0.08 33 1.55 ± 0.11 1.20 ± 0.14 1.52 ± 0.10 0.95 ± 0.06 1.37 ± 0.16 0.96 ± 0.08 36 1.39 ± 0.06 1.68 ± 0.24 1.25 ± 0.20 0.75 ± 0.14 1.36 ± 0.14 0.97 ± 0.46 42 1.58 ± 0.28 d 2.45 ± 0.29 1.84 ± 0.35 1.05 ± 0.07 1.25 ± 0.08 d 0.91 ± 0.28 48 1.71 ± 0.21 1.64 ± 0.25 1.89 ± 0.13 1.35 ± 0.06 1.70 ± 0.17 1.82 ± 0.48 Arginine 30 0.20 ± 0.01 0.16 ± 0.05 0.16 ± 0.05 0.11 ± 0.01 0.09 ± 0.00 0.09 ± 0.00 33 0.16 ± 0.03 0.15 ± 0.01 0.16 ± 0.02 0.11 ± 0.01 0.09 ± 0.01 0.12 ± 0.03 36 0.14 ± 0.05 0.16 ± 0.04 0.12 ± 0.01 0.12 ± 0.01 0.18 ± 0.02 0.13 ± 0.06 42 0.16 ± 0.03 0.19 ± 0.04 0.20 ± 0.03 0.14 ± 0.03 0.11 ± 0.03 0.09 ± 0.01 48 0.24 ± 0.08 0.16 ± 0.04 0.25 ± 0.03 0.14 ± 0.01 0.25 ± 0.09 0.18 ± 0.06 Asprgine 30 4.93 ± 0.10 3.68 ± 1.24 3.68 ± 1.24 1.58 ± 0.37 12.07 ± 3.68 12.07 ± 3.68 33 5.10 ± 0.35 3.84 ± 0.18 4.98 ± 0.51 1.11 ± 0.19 12.29 ± 1.81 5.15 ± 1.59 36 4.92 ± 1.29 3.46 ± 0.66 d 4.77 ± 0.79 1.53 ± 0.18 21.95 ± 5.31 3.13 ± 0.16 d 42 3.45 ± 1.40 3.93 ± 0.51 11.04 ± 0.38 3.42 ± 0.37 15.37 ± 2.03 3.24 ± 0.46 48 15.07 ± 3.54 3.94 ± 0.49 19.25 ± 1.77 4.13 ± 0.74 9.09 ± 3.06 3.91 ± 1.30 Asprtte 30 0.85 ± 0.05 0.76 ± 0.25 0.76 ± 0.25 0.55 ± 0.14 0.82 ± 0.10 0.82 ± 0.10 33 1.07 ± 0.06 0.79 ± 0.06 0.92 ± 0.08 0.48 ± 0.07 0.81 ± 0.08 0.57 ± 0.04 36 0.66 ± 0.14 0.90 ± 0.17 0.97 ± 0.44 0.67 ± 0.07 0.85 ± 0.19 0.75 ± 0.24 42 0.76 ± 0.24 0.99 ± 0.10 0.68 ± 0.08 0.70 ± 0.10 0.73 ± 0.08 0.45 ± 0.06 48 0.83 ± 0.15 0.69 ± 0.15 0.72 ± 0.10 0.89 ± 0.11 1.28 ± 0.13 0.72 ± 0.22 Glutmte 30 4.16 ± 0.24 3.70 ± 0.48 3.70 ± 0.48 2.24 ± 0.13 2.44 ± 0.11 2.44 ± 0.11 33 3.26 ± 0.23 3.26 ± 0.24 3.52 ± 0.44 2.65 ± 0.05 2.69 ± 0.28 d 2.15 ± 0.31 d 36 2.78 ± 0.62 3.81 ± 0.46 3.08 ± 0.55 2.20 ± 0.10 2.98 ± 0.37 2.06 ± 0.19 42 3.16 ± 0.33 3.76 ± 0.36 3.16 ± 0.41 2.41 ± 0.07 2.28 ± 0.21 1.82 ± 0.58 48 3.00 ± 0.33 2.48 ± 0.27 3.31 ± 0.09 1.95 ± 0.09 1.74 ± 0.36 2.05 ± 0.40 Glutmine 30 8.90 ± 0.18 10.46 ± 2.11 10.46 ± 2.11 1.16 ± 0.89 7.12 ± 0.36 d 7.12 ± 0.36 d 33 7.29 ± 0.41 11.12 ± 0.47 12.02 ± 1.50 2.00 ± 0.37 10.37 ± 1.05 6.27 ± 1.73 36 6.96 ± 2.86 11.73 ± 0.59 6.17 ± 0.20 2.35 ± 0.12 12.92 ± 1.48 4.50 ± 2.34 42 8.06 ± 1.29 14.36 ± 1.87 8.97 ± 1.13 2.21 ± 0.26 10.99 ± 3.63 2.43 ± 0.65 48 7.48 ± 2.09 10.40 ± 0.82 5.26 ± 0.34 4.29 ± 1.01 8.16 ± 1.74 5.59 ± 1.29 Glyine 30 0.13 ± 0.01 0.14 ± 0.04 0.14 ± 0.04 0.10 ± 0.02 0.30 ± 0.01 0.30 ± 0.01 33 0.10 ± 0.00 0.12 ± 0.01 0.11 ± 0.01 0.10 ± 0.00 0.34 ± 0.00 0.29 ± 0.10 36 0.10 ± 0.02 0.13 ± 0.03 0.09 ± 0.01 0.09 ± 0.01 0.26 ± 0.03 0.27 ± 0.09 42 0.10 ± 0.01 0.15 ± 0.01 0.12 ± 0.03 0.14 ± 0.03 0.26 ± 0.05 0.22 ± 0.01 48 0.12 ± 0.01 0.11 ± 0.02 0.11 ± 0.01 0.15 ± 0.03 0.21 ± 0.10 0.21 ± 0.01 Histidine 30 0.49 ± 0.05 0.49 ± 0.15 0.49 ± 0.15 0.47 ± 0.03 1.66 ± 0.08 1.66 ± 0.08 33 0.46 ± 0.03 0.47 ± 0.02 0.47 ± 0.07 0.45 ± 0.02 1.79 ± 0.12 0.87 ± 0.33 36 0.47 ± 0.10 0.50 ± 0.04 0.52 ± 0.07 0.49 ± 0.03 1.25 ± 0.13 0.83 ± 0.23 42 0.51 ± 0.07 0.65 ± 0.08 0.68 ± 0.10 0.56 ± 0.03 1.27 ± 0.18 0.57 ± 0.01 48 1.06 ± 0.11 0.83 ± 0.02 0.90 ± 0.06 0.73 ± 0.08 0.84 ± 0.12 0.75 ± 0.11 Isoleuine 30 0.16 ± 0.02 0.21 ± 0.05 0.21 ± 0.05 0.11 ± 0.01 0.55 ± 0.01 0.55 ± 0.01 33 0.15 ± 0.02 0.18 ± 0.01 0.16 ± 0.03 0.09 ± 0.00 0.59 ± 0.04 0.27 ± 0.13 36 0.15 ± 0.02 0.18 ± 0.02 0.19 ± 0.03 0.11 ± 0.01 0.38 ± 0.02 0.22 ± 0.09 42 0.17 ± 0.02 0.29 ± 0.02 0.19 ± 0.02 0.18 ± 0.06 0.42 ± 0.11 0.10 ± 0.03 48 0.41 ± 0.07 0.42 ± 0.12 0.26 ± 0.02 0.16 ± 0.02 0.26 ± 0.03 0.12 ± 0.06 Leuine 30 0.13 ± 0.01 0.19 ± 0.05 0.19 ± 0.05 0.11 ± 0.01 0.54 ± 0.10 0.54 ± 0.10 33 0.13 ± 0.01 0.17 ± 0.01 0.14 ± 0.03 0.09 ± 0.00 0.44 ± 0.03 0.22 ± 0.07 36 0.13 ± 0.02 0.17 ± 0.02 0.17 ± 0.02 0.12 ± 0.01 0.35 ± 0.02 0.18 ± 0.07 42 0.14 ± 0.01 0.25 ± 0.02 0.16 ± 0.02 0.16 ± 0.05 0.40 ± 0.10 0.09 ± 0.02 d 48 0.31 ± 0.07 0.32 ± 0.12 0.19 ± 0.02 0.16 ± 0.02 0.29 ± 0.03 0.21 ± 0.06 Lysine 30 0.19 ± 0.01 0.21 ± 0.04 0.21 ± 0.04 0.17 ± 0.08 0.38 ± 0.07 0.38 ± 0.07 33 0.16 ± 0.02 0.14 ± 0.03 0.15 ± 0.01 0.15 ± 0.07 0.27 ± 0.02 0.19 ± 0.03 36 0.14 ± 0.02 0.15 ± 0.01 0.19 ± 0.02 0.17 ± 0.00 0.26 ± 0.02 0.18 ± 0.05 42 0.14 ± 0.00 0.17 ± 0.01 0.16 ± 0.01 0.17 ± 0.01 0.31 ± 0.04 0.15 ± 0.02 48 0.21 ± 0.01 0.19 ± 0.02 0.20 ± 0.02 0.20 ± 0.01 0.34 ± 0.01 0.22 ± 0.06

Hordeum mritimum Hordeum vulgre Control Sl stress Slt removl Control Slt stress Dy Slt removl Methionine 30 0.07 ± 0.00 0.07 ± 0.02 0.07 ± 0.02 0.05 ± 0.00 0.11 ± 0.01 0.11 ± 0.01 33 0.06 ± 0.00 0.07 ± 0.00 0.07 ± 0.01 0.06 ± 0.00 0.12 ± 0.01 0.07 ± 0.01 36 0.06 ± 0.02 0.07 ± 0.01 0.07 ± 0.01 0.06 ± 0.00 0.08 ± 0.00 0.07 ± 0.03 42 0.07 ± 0.00 0.09 ± 0.01 0.07 ± 0.01 0.08 ± 0.03 0.12 ± 0.03 0.06 ± 0.01 48 0.07 ± 0.01 0.06 ± 0.01 0.06 ± 0.00 0.10 ± 0.00 0.13 ± 0.01 0.09 ± 0.03 Ornithine 30 0.55 ± 0.03 0.68 ± 0.05 0.68 ± 0.05 0.18 ± 0.05 0.39 ± 0.02 d 0.39 ± 0.02 d 33 0.51 ± 0.03 0.36 ± 0.01 0.54 ± 0.08 0.18 ± 0.07 0.40 ± 0.03 d 0.31 ± 0.04 36 0.48 ± 0.05 0.45 ± 0.13 0.79 ± 0.22 0.14 ± 0.00 0.32 ± 0.02 d 0.26 ± 0.11 d 42 0.43 ± 0.05 0.57 ± 0.02 0.41 ± 0.05 0.26 ± 0.02 0.37 ± 0.05 0.19 ± 0.05 48 0.43 ± 0.03 0.46 ± 0.13 0.46 ± 0.01 0.16 ± 0.01 0.37 ± 0.01 0.18 ± 0.04 Phenyllnine 30 0.22 ± 0.01 0.34 ± 0.10 0.34 ± 0.10 0.18 ± 0.02 0.96 ± 0.04 0.96 ± 0.04 33 0.26 ± 0.02 0.31 ± 0.01 0.30 ± 0.04 0.16 ± 0.01 1.11 ± 0.11 d 0.53 ± 0.22 36 0.24 ± 0.05 0.32 ± 0.03 0.30 ± 0.04 0.20 ± 0.01 0.55 ± 0.06 0.45 ± 0.21 42 0.28 ± 0.03 0.52 ± 0.04 0.33 ± 0.04 0.31 ± 0.06 0.77 ± 0.16 0.20 ± 0.05 48 0.65 ± 0.11 0.62 ± 0.18 0.45 ± 0.04 0.34 ± 0.04 0.75 ± 0.07 0.55 ± 0.18 Proline 30 2.39 ± 0.06 8.48 ± 0.26 8.48 ± 0.26 1.9 ± 0.1 20.6 ± 3.3 d 20.6 ± 3.3 d 33 2.11 ± 0.18 5.85 ± 0.43 4.39 ± 0.356 1.8 ± 0.0 27.1 ± 1.4 d 7.8 ± 0.1 e 36 2.80 ± 0.41 7.31 ± 0.49 4.43 ± 0.844 1.9 ± 0.1 d 26.5 ± 0.3 e 7.5 ± 1.5 42 2.95 ± 0.08 13.93 ± 0.43 3.53 ± 0.767 1.9 ± 0.0 19.8 ± 0.4 d 2.6 ± 0.3 48 3.61 ± 1.12 15.19 ± 2.14 2.31 ± 0.248 1.9 ± 0.1 14.8 ± 1.7 2.2 ± 0.3 Serine 30 2.11 ± 0.13 1.71 ± 0.45 1.71 ± 0.45 0.77 ± 0.26 2.63 ± 0.11 2.63 ± 0.11 33 1.85 ± 0.12 1.73 ± 0.11 2.32 ± 0.17 1.12 ± 0.19 2.57 ± 0.22 1.42 ± 0.07 36 1.72 ± 0.34 2.08 ± 0.53 1.74 ± 0.27 0.93 ± 0.05 3.63 ± 0.37 1.11 ± 0.33 42 1.77 ± 0.38 2.49 ± 0.31 2.31 ± 0.37 0.98 ± 0.12 2.31 ± 0.03 0.78 ± 0.21 48 2.13 ± 0.17 1.85 ± 0.21 2.26 ± 0.15 1.20 ± 0.07 2.58 ± 0.43 1.29 ± 0.48 Threonine 30 0.64 ± 0.04 0.48 ± 0.14 0.48 ± 0.14 0.30 ± 0.02 0.61 ± 0.05 0.61 ± 0.05 33 0.57 ± 0.04 0.42 ± 0.03 0.49 ± 0.06 0.35 ± 0.00 0.53 ± 0.02 0.39 ± 0.04 36 0.46 ± 0.09 0.56 ± 0.09 0.44 ± 0.06 0.33 ± 0.01 0.70 ± 0.07 0.43 ± 0.21 42 0.52 ± 0.07 0.65 ± 0.07 0.57 ± 0.09 0.48 ± 0.10 0.60 ± 0.05 0.28 ± 0.06 48 0.70 ± 0.09 0.61 ± 0.01 0.69 ± 0.05 0.53 ± 0.05 0.54 ± 0.24 0.70 ± 0.19 Tryptophn 30 0.15 ± 0.04 0.13 ± 0.02 0.13 ± 0.02 0.16 ± 0.01 0.28 ± 0.03 0.28 ± 0.03 33 0.12 ± 0.01 0.12 ± 0.01 0.12 ± 0.01 0.17 ± 0.01 0.36 ± 0.02 0.23 ± 0.03 36 0.11 ± 0.02 0.13 ± 0.01 0.12 ± 0.01 0.17 ± 0.02 0.21 ± 0.02 0.22 ± 0.07 42 0.12 ± 0.01 0.18 ± 0.01 0.12 ± 0.02 0.19 ± 0.01 0.26 ± 0.04 0.17 ± 0.02 48 0.17 ± 0.03 0.15 ± 0.03 0.14 ± 0.00 0.22 ± 0.01 0.27 ± 0.02 0.21 ± 0.06 Tyrosine 30 0.47 ± 0.03 0.46 ± 0.14 0.46 ± 0.14 0.20 ± 0.02 0.57 ± 0.08 0.57 ± 0.08 33 0.53 ± 0.03 0.59 ± 0.07 0.81 ± 0.12 0.22 ± 0.00 0.66 ± 0.07 0.39 ± 0.08 d 36 0.59 ± 0.13 0.78 ± 0.14 0.61 ± 0.08 0.34 ± 0.03 0.48 ± 0.04 0.43 ± 0.13 42 0.67 ± 0.11 1.18 ± 0.13 0.87 ± 0.10 0.50 ± 0.09 d 0.64 ± 0.12 0.40 ± 0.14 d 48 0.74 ± 0.17 0.73 ± 0.18 0.50 ± 0.04 1.17 ± 0.02 1.07 ± 0.08 0.99 ± 0.24 Vline 30 0.16 ± 0.01 0.20 ± 0.05 0.20 ± 0.05 0.12 ± 0.01 0.58 ± 0.06 0.58 ± 0.06 33 0.15 ± 0.01 0.18 ± 0.03 0.17 ± 0.02 0.12 ± 0.01 0.76 ± 0.04 0.32 ± 0.16 d 36 0.14 ± 0.02 0.19 ± 0.03 0.18 ± 0.02 0.16 ± 0.04 0.48 ± 0.02 0.28 ± 0.04 42 0.15 ± 0.01 0.28 ± 0.02 0.18 ± 0.02 0.18 ± 0.05 0.41 ± 0.10 0.13 ± 0.02 48 0.36 ± 0.16 0.36 ± 0.12 0.18 ± 0.04 0.17 ± 0.01 0.32 ± 0.03 0.22 ± 0.06

Tle S2. Sodium, hloride nd potssium ontent of entire shoot t 30 nd 48 dys fter sowing (DAS), older lef tissues lredy present efore 30 DAS nd still present t 48 DAS nd lef younger tissues developed in the period 30-48 DAS fter the esstion of slinity tretment. H. mritimum nd H. vulgre plnts of seond independent experiment run with sme experimentl design s in Tle S1 nd Figure 1. Hrvests were done t 30 nd 48 DAS. Vlues re men s.d. (n = 3) Hordeum mritimum Hordeum vulgre Dy Control Sl stress Slt removl Control Slt stress Slt removl Chloride (mmol g 1 DW) 30 0.45 ± 0.02 1.00 ± 0.12 1.00 ± 0.12 0.79 ± 0.01 1.93 ± 0.07 d 1.93 ± 0.07 d 33 0.50 ± 0.01 1.08 ± 0.18 0.76 ± 0.03 0.76 ± 0.04 2.09 ± 0.03 d 1.61 ± 0.02 e 36 0.48 ± 0.01 1.29 ± 0.11 0.66 ± 0.04 0.68 ± 0.04 1.92 ± 0.03 d 1.70 ± 0.12 e 42 0.63 ± 0.02 1.80 ± 0.08 0.64 ± 0.11 0.66 ± 0.03 1.71 ± 0.04 1.23 ± 0.07 48 0.60 ± 0.03 1.62 ± 0.35 0.50 ± 0.07 0.78 ± 0.15 1.71 ± 0.10 1.03 ± 0.03 d Nitrte (mmol g 1 DW) 30 0.69 ± 0.04 0.24 ± 0.00 0.24 ± 0.00 0.85 ± 0.05 0.17 ± 0.01 d 0.17 ± 0.01 d 33 0.78 ± 0.05 0.23 ± 0.00 0.23 ± 0.03 0.77 ± 0.05 0.14 ± 0.01 0.17 ± 0.04 36 0.77 ± 0.02 0.27 ± 0.03 0.30 ± 0.06 0.71 ± 0.06 0.12 ± 0.01 0.25 ± 0.02 42 1.02 ± 0.08 0.32 ± 0.00 0.57 ± 0.08 0.69 ± 0.05 0.11 ± 0.01 d 0.60 ± 0.04 48 0.79 ± 0.05 0.21 ± 0.02 0.55 ± 0.02 0.69 ± 0.10 0.09 ± 0.00 d 0.70 ± 0.05 Potssium (mmol g 1 DW) 30 0.44 ± 0.01 0.35 ± 0.03 0.35 ± 0.03 0.79 ± 0.02 0.42 ± 0.01 d 0.42 ± 0.01 d 33 0.51 ± 0.01 0.34 ± 0.04 0.31 ± 0.02 0.58 ± 0.03 0.38 ± 0.01 d 0.41 ± 0.02 d 36 0.49 ± 0.04 0.31 ± 0.03 0.37 ± 0.03 0.61 ± 0.04 0.32 ± 0.04 0.48 ± 0.01 42 0.61 ± 0.04 0.36 ± 0.01 0.55 ± 0.04 0.59 ± 0.02 0.21 ± 0.02 0.55 ± 0.03 48 0.46 ± 0.06 0.28 ± 0.03 0.50 ± 0.01 0.61 ± 0.03 0.20 ± 0.01 d 0.55 ± 0.04 Sodium (mmol g 1 DW) 30 0.14 ± 0.02 0.40 ± 0.05 0.40 ± 0.05 0.14 ± 0.05 1.02 ± 0.07 1.02 ± 0.07 33 0.16 ± 0.04 0.42 ± 0.02 0.38 ± 0.03 0.13 ± 0.02 1.07 ± 0.04 0.99 ± 0.07 36 0.18 ± 0.03 0.43 ± 0.06 0.34 ± 0.04 0.13 ± 0.02 1.21 ± 0.11 0.95 ± 0.06 42 0.20 ± 0.05 0.68 ± 0.03 0.30 ± 0.05 0.14 ± 0.02 1.10 ± 0.10 d 0.82 ± 0.05 e 48 0.20 ± 0.02 0.63 ± 0.02 0.20 ± 0.02 0.15 ± 0.01 1.45 ± 0.15 d 0.62 ± 0.08 Potssium:Sodium 30 3.23 ± 0.39 0.88 ± 0.10 0.88 ± 0.11 5.56 ± 0.70 0.41 ± 0.05 d 0.41 ± 0.05 d 33 3.28 ± 0.40 0.81 ± 0.10 0.82 ± 0.10 4.35 ± 0.55 0.35 ± 0.05 d 0.41 ± 0.05 d 36 2.72 ± 0.22 0.74 ± 0.09 1.09 ± 0.11 4.62 ± 0.61 d 0.26 ± 0.03 e 0.51 ± 0.06 f 42 3.02 ± 0.40 0.52 ± 0.05 1.81 ± 0.19 4.36 ± 0.46 d 0.19 ± 0.03 e 0.68 ± 0.08 f 48 2.34 ± 0.33 0.44 ± 0.04 2.50 ± 0.32 4.09 ± 0.50 0.14 ± 0.02 d 0.89 ± 0.10 e

Tle S3. Sodium, hloride nd potssium ontent of entire shoot t 30 nd 48 dys fter sowing (DAS), older lef tissues lredy present efore 30 DAS nd still present t 48 DAS nd lef younger tissues developed in the period 30-48 DAS fter the esstion of slinity tretment nd expressed on fresh nd dry weight ses The H. mritimum nd H. vulgre plnt mteril used ws from seond independent experiment run with sme experimentl design s in Tle S1 nd Figure 1. Hrvests were done t 30 nd 48 DAS. Vlues re men s.d. (n = 3) Dy Chloride (μmol g 1 FW) 30 155 ± 12.5 242 ± 22.3 48 66.3 ± 10.0 100 ± 14.3 54.2 ± 9.9 100 ± 12.6 191 ± 17.8 60.7 ± 3.5 Sodium (μmol g 1 FW) 30 59.8 ± 8.7 126 ± 14.2 48 21.3 ± 5.4 25.6 ± 5.4 19.8 ± 1.2 54.9 ± 10.7 94.7 ± 26.3 37.8 ± 8.3 Potssium (μmol g 1 FW) 30 48.5 ± 7.3 60.4 ± 8.6 48 59.4 ± 4.8 39.2 ± 4.8 66.6 ± 3.2 62.4 ± 4.9 51.3 ± 1.3 67.2 ± 2.8 Chloride (mmol g 1 DW) Hordeum mritimum 30 1.09 ± 0.09 3.00 ± 0.28 48 0.54 ± 0.08 0.94 ± 0.13 0.51 ± 0.09 1.10 ± 0.14 1.55 ± 0.14 0.91 ± 0.05 Sodium (mmol g 1 DW) 30 0.42 ± 0.06 1.56 ± 0.18 48 0.17 ± 0.04 0.24 ± 0.05 0.19 ± 0.01 0.61 ± 0.12 0.77 ± 0.21 0.57 ± 0.12 Potssium (mmol g 1 DW) 30 0.34 ± 0.05 0.75 ± 0.11 Hordeum vulgre Entire shoot Older tissue Younger tissue Entire shoot Older tissue Younger tissue 48 0.49 ± 0.04 0.37 ± 0.05 0.63 ± 0.03 0.69 ± 0.05 0.42 ± 0.01 1.01 ± 0.04

Fig. S1. Photogrphs showing representtive H. vulgre plnts t 30, 36, 42, 48 DAS. Fig. S2. Photogrphs showing representtive H. mritimum plnts t 30, 36, 48 DAS.

Fig. S3. Shoot fresh (A) nd dry (B) weights of H. mritimum (Hm) nd H. vulgre (Hv) plnts under ontrol, stress (200 mm NCl) nd slt removl tretments. Slt ws grdully dded to slinity tretments strting from 15 DAS. Slt removl tretment strted from 30 DAS. Hrvest ws done t 30, 33, 36, 42 nd 48 DAS. Metolites were expressed s µmol g -1 FW. Vlues re men s.d. (n=3).