Metabolomic analysis revealed differential adaptation to salinity and alkalinitystress in Kentucky bluegrass(Poa pratensis)
Longxing Hu, Pingping Zhang, Ying Jiang, Jinmin Fu*
Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, The Chinese Academy of Sciences, Wuhan, Hubei 430074, PR China
*Corresponding author:
Supplementary materials
Table S1 Preparation of the treatment solutions (control, salinity, and alkaline solutions(carbonate -bicarbonate)
Treatment / pH(25°C) / Na+ concentration
(mM) / 50 mM NaHCO3 (%) / 50 mM Na2CO3 (%)
Control / 5.33 / 0 / 0 / 0
Salinity / 5.41 / 75 / 0 / 0
Alkalinity / 9.50 / 75 / 50 / 50
Table S2 Overview of the metabolite reporting list (See in a separate excel file)
Table S3Response ratios (relative to the internal standard ribitol) of identified metabolites in leaves of Kentucky bluegrass under salinity and alkalinity stress. (MC, ‘Midnight’ control; MS, ‘Midnight’ salinity; MA, ‘Midnight’ alkalinity; VC, ‘Voyager II’ control; VS, ‘Voyager II’ salinity; VA, ‘Voyager II’ alkalinity)
Category / label / MC / MS / MA / VC / VS / VAAmino acids / Valine / 0.0827 / 0.1530 / 0.1349 / 0.1014 / 0.1267 / 0.1030
Alanine / 0.2339 / 0.3147 / 0.1545 / 0.2151 / 0.2132 / 0.0652
Proline / 0.0100 / 0.4822 / 0.3051 / 0.0100 / 0.5976 / 0.3034
Serine / 1.0284 / 2.2495 / 0.4177 / 0.7385 / 2.1034 / 0.2445
Leucine / 0.0335 / 0.0913 / 0.1007 / 0.0170 / 0.0765 / 0.0372
Threonine / 1.5837 / 2.1566 / 0.5768 / 1.5210 / 3.0164 / 0.3252
Glycine / 0.6378 / 0.6684 / 0.6047 / 0.6426 / 0.7388 / 0.6068
Methionine / 0.0162 / 0.0209 / 0.0203 / 0.0084 / 0.0110 / 0.0131
Aspartate / 0.8386 / 1.0495 / 0.4515 / 0.3726 / 1.2671 / 0.2400
beta-Alanine / 0.0297 / 0.0273 / 0.0388 / 0.0305 / 0.0229 / 0.0131
Homoserine / 0.0053 / 0.0091 / 0.0001 / 0.0054 / 0.0101 / 0.0002
5-Oxoproline / 0.8105 / 2.3133 / 0.5088 / 1.0564 / 2.0564 / 0.4974
Glutamate / 0.3485 / 0.7539 / 0.2759 / 0.3684 / 0.5195 / 0.2855
Phenylalanine / 0.0512 / 0.0665 / 0.0386 / 0.0584 / 0.0628 / 0.0405
Asparagine / 0.1691 / 0.4967 / 0.0430 / 0.1501 / 0.3325 / 0.0101
Glutamine / 0.5850 / 1.5787 / 0.1498 / 0.3625 / 0.6333 / 0.0369
Lysine / 0.0576 / 0.1313 / 0.0289 / 0.0234 / 0.0418 / 0.0011
Organic acids / Oxalate / 0.4231 / 0.2277 / 0.3926 / 0.5430 / 0.1813 / 0.8624
Phosphate / 0.4425 / 0.4756 / 0.4134 / 0.3819 / 0.4929 / 0.2781
Malonate / 0.0301 / 0.0433 / 0.0631 / 0.0324 / 0.0577 / 0.0383
Maleiate / 0.3779 / 0.1950 / 0.5440 / 0.3151 / 0.1200 / 0.4478
Succinate / 0.2343 / 0.1950 / 0.4651 / 0.4593 / 0.3251 / 0.6774
Glycerate / 0.5172 / 0.4982 / 0.8157 / 0.3385 / 0.3360 / 0.5097
Malate / 18.9359 / 14.9959 / 26.6737 / 15.5715 / 13.2323 / 30.4370
Butanoate / 6.2954 / 5.6705 / 3.8979 / 4.7905 / 4.2863 / 2.1748
2-Oxoglutarate / 0.5943 / 0.2542 / 1.8614 / 1.3932 / 1.2544 / 7.7642
Threonate / 0.0823 / 0.0562 / 0.1052 / 0.0878 / 0.0446 / 0.0402
Aconitate / 0.0377 / 0.0466 / 0.0912 / 0.0865 / 0.0399 / 0.0986
Shikimate / 1.2851 / 0.7456 / 1.7623 / 2.6716 / 1.0593 / 3.0916
Citrate / 8.1770 / 5.1259 / 13.0760 / 8.3135 / 6.6149 / 13.1235
Isocitrate / 5.5418 / 0.9108 / 9.6854 / 5.4562 / 0.9759 / 8.2418
Gluconate / 0.0733 / 0.0676 / 0.0714 / 0.1006 / 0.0894 / 0.1279
Glucarate / 0.7323 / 0.4813 / 1.5736 / 0.5934 / 0.5994 / 0.8711
Galactarate / 0.1784 / 0.0873 / 0.2980 / 0.1118 / 0.0936 / 0.1828
Sugars / Glycerol / 3.5754 / 5.2073 / 3.7067 / 2.8947 / 1.7384 / 3.4263
Fructose / 26.3715 / 22.7787 / 16.8706 / 32.9870 / 23.8916 / 12.2238
Glucose / 16.7738 / 15.7537 / 7.3536 / 17.6631 / 15.0257 / 5.1873
Mannose / 4.9703 / 4.4879 / 4.8559 / 6.6081 / 6.4476 / 9.1875
Myo-Inositol / 0.7634 / 0.6597 / 0.7540 / 0.7010 / 0.6682 / 0.6984
Sucrose / 20.7293 / 25.1596 / 25.6823 / 17.3251 / 20.8832 / 20.0217
Trehalose / 0.6260 / 1.4745 / 0.3881 / 0.5702 / 0.8378 / 0.2800
Gentiobiose / 0.7794 / 0.6443 / 0.3602 / 1.1848 / 1.6771 / 0.6916
Galactinol / 0.2843 / 0.2832 / 0.3008 / 0.1027 / 0.1369 / 0.0964
Fatty acids / Palmitic acid / 0.1825 / 0.1711 / 0.2322 / 0.2179 / 0.2781 / 0.3810
Linoleic acid / 0.0455 / 0.0432 / 0.0482 / 0.0490 / 0.0273 / 0.0397
Linolenic acid / 0.2716 / 0.2147 / 0.1704 / 0.2820 / 0.1751 / 0.1283
Stearic acid / 0.1459 / 0.1266 / 0.1434 / 0.1682 / 0.1892 / 0.2151
Others / Cinnamic acid / 0.1470 / 0.1552 / 0.1959 / 0.2030 / 0.2130 / 0.2613
Cinnamate / 0.2673 / 0.2334 / 0.5653 / 0.1097 / 0.1589 / 0.0862
Tocopherol / 0.0124 / 0.0120 / 0.0072 / 0.0169 / 0.0164 / 0.0285
Sitosterol / 0.1122 / 0.0989 / 0.1077 / 0.1296 / 0.1117 / 0.1442
*Identifications based on TMS and TBS derivatized samples are combined.
Table S4 Variation of metabolites in leaves of Kentucky bluegrass between treatment (salinity, alkalinity) and the control, and between salinity and alkalinity using two sample t-tests at p < 0.05(MC, ‘Midnight’ control; MS, ‘Midnight’ salinity; MA, ‘Midnight’ alkalinity; VC, ‘Voyager II’ control; VS, ‘Voyager II’ salinity; VA, ‘Voyager II’ alkalinity)
Relative to the control / higher in MS / higher in VS / higher in MACategory / Identified / Variations / MS / MA / VS / VA / MS vs. MA / MS vs. VS / VS vs. VA / MA vs. VA
Amino acids / 17 / Increasing / 15 / 5 / 12 / 3 / 12 / 9 / 14 / 6
Decreasing / 0 / 11 / 1 / 12
Organic acids / 17 / Increasing / 1 / 13 / 2 / 11 / 1 / 3 / 3 / 6
Decreasing / 10 / 1 / 9 / 3
Sugars / 9 / Increasing / 2 / 1 / 3 / 1 / 5 / 2 / 5 / 0
Decreasing / 2 / 4 / 2 / 4
Others / 8 / Increasing / 0 / 3 / 2 / 4 / 1 / 0 / 1 / 2
Decreasing / 2 / 2 / 3 / 3
total / 51
Table S5 Metabolites changes in salinity- or alkalinity-treated leaves of Kentucky bluegrass relative to the control. (MC, ‘Midnight’ control; MS, ‘Midnight’ salinity; MA, ‘Midnight’ alkalinity; VC, ‘Voyager II’ control; VS, ‘Voyager II’ salinity; VA, ‘Voyager II’ alkalinity)
Category / label / MC / MS / MA / VC / VS / VAAmino acids / Valine / 1 / 1.851 / 1.632 / 1 / 1.250 / 1.015
Alanine / 1 / 1.345 / 0.660 / 1 / 0.991 / 0.303
Proline / 1 / 48.219 / 30.510 / 1 / 59.762 / 30.343
Serine / 1 / 2.187 / 0.406 / 1 / 2.848 / 0.331
Leucine / 1 / 2.728 / 3.008 / 1 / 4.500 / 2.189
Threonine / 1 / 1.362 / 0.364 / 1 / 1.983 / 0.214
Glycine / 1 / 1.048 / 0.948 / 1 / 1.150 / 0.944
Methionine / 1 / 1.287 / 1.250 / 1 / 1.308 / 1.556
Aspartate / 1 / 1.251 / 0.538 / 1 / 3.401 / 0.644
beta-Alanine / 1 / 0.920 / 1.307 / 1 / 0.750 / 0.431
Homoserine / 1 / 1.715 / 0.022 / 1 / 1.889 / 0.031
5-Oxoproline / 1 / 2.854 / 0.628 / 1 / 1.947 / 0.471
Glutamate / 1 / 2.163 / 0.792 / 1 / 1.410 / 0.775
Phenylalanine / 1 / 1.298 / 0.753 / 1 / 1.075 / 0.694
Asparagine / 1 / 2.937 / 0.254 / 1 / 2.215 / 0.067
Glutamine / 1 / 2.699 / 0.256 / 1 / 1.747 / 0.102
Lysine / 1 / 2.278 / 0.502 / 1 / 1.786 / 0.048
Organic acids / Oxalate / 1 / 0.538 / 0.928 / 1 / 0.334 / 1.588
Phosphate / 1 / 1.075 / 0.934 / 1 / 1.291 / 0.728
Malonate / 1 / 1.437 / 2.092 / 1 / 1.781 / 1.181
Maleiate / 1 / 0.516 / 1.440 / 1 / 0.381 / 1.421
Succinate / 1 / 0.832 / 1.985 / 1 / 0.708 / 1.475
Glycerate / 1 / 0.963 / 1.577 / 1 / 0.993 / 1.506
Malate / 1 / 0.792 / 1.409 / 1 / 0.850 / 1.955
Butanoate / 1 / 0.901 / 0.619 / 1 / 0.895 / 0.454
2-Oxoglutarate / 1 / 0.428 / 3.132 / 1 / 0.900 / 5.573
Threonate / 1 / 0.682 / 1.278 / 1 / 0.508 / 0.457
Aconitate / 1 / 1.234 / 2.415 / 1 / 0.461 / 1.139
Shikimate / 1 / 0.580 / 1.371 / 1 / 0.397 / 1.157
Citrate / 1 / 0.627 / 1.599 / 1 / 0.796 / 1.579
Isocitrate / 1 / 0.164 / 1.748 / 1 / 0.179 / 1.511
Gluconate / 1 / 0.922 / 0.974 / 1 / 0.888 / 1.271
Glucarate / 1 / 0.657 / 2.149 / 1 / 1.010 / 1.468
Galactarate / 1 / 0.489 / 1.670 / 1 / 0.838 / 1.636
Sugars / Glycerol / 1 / 1.456 / 1.037 / 1 / 0.601 / 1.184
Fructose / 1 / 0.864 / 0.640 / 1 / 0.724 / 0.371
Glucose / 1 / 0.939 / 0.438 / 1 / 0.851 / 0.294
Mannose / 1 / 0.903 / 0.977 / 1 / 0.976 / 1.390
Myo-Inositol / 1 / 0.864 / 0.988 / 1 / 0.953 / 0.996
Sucrose / 1 / 1.214 / 1.239 / 1 / 1.205 / 1.156
Trehalose / 1 / 2.355 / 0.620 / 1 / 1.469 / 0.491
Gentiobiose / 1 / 0.827 / 0.462 / 1 / 1.416 / 0.584
Galactinol / 1 / 0.996 / 1.058 / 1 / 1.333 / 0.938
Fatty acids / Palmitic acid / 1 / 0.938 / 1.273 / 1 / 1.276 / 1.749
Linoleic acid / 1 / 0.950 / 1.060 / 1 / 0.558 / 0.811
Linolenic acid / 1 / 0.790 / 0.627 / 1 / 0.621 / 0.455
Stearic acid / 1 / 0.868 / 0.983 / 1 / 1.125 / 1.279
Others / Cinnamic acid / 1 / 1.055 / 1.332 / 1 / 1.049 / 1.287
Cinnamate / 1 / 0.873 / 2.115 / 1 / 1.449 / 0.786
Tocopherol / 1 / 0.970 / 0.578 / 1 / 0.972 / 1.687
Sitosterol / 1 / 0.881 / 0.960 / 1 / 0.861 / 1.112
Fig.S1The metabolic differences between salinity and alkalinity stress in ‘Midnight’ (a) and ‘Voyager II’ (b). The selected metabolites have variations >50% and/or significant differences according to the two sample t-tests (p < 0.05, n = 4) were presented as logarithmic ratios after normalized to the respective controls (MC, ‘Midnight’ control; MS, ‘Midnight’ salt; MA, ‘Midnight’ alkali; VC, ‘Voyager II’ control; VS, ‘Voyager II’ salt; VA, ‘Voyager II’ alkali) sampled at 7 d of treatment.
Fig. S2The metabolic differences between ‘Midnight’ and ‘Voyager II’ under salinity (a) or alkalinity (b) stress. The selected metabolites have variations >50% and/or significant differences according to the two sample t-tests (p < 0.05, n = 4) were presented as logarithmic ratios after normalized to the respective controls (MC, ‘Midnight’ control; MS, ‘Midnight’ salt; MA, ‘Midnight’ alkali; VC, ‘Voyager II’ control; VS, ‘Voyager II’ salt; VA, ‘Voyager II’ alkali) sampled at 7 d of treatment.
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