Supplementary materials
Table S1 Differential metabolites between transgenic backcross breeding seeds and its non-transgenic parent
Metabolite / PubChem CID / Identification Level a / Retention Time (min) / RI Index / Related pathways / Quantification Ions [m/z] / Fold-change b(8Km732 /MH3301)
β-alanine / 239 / 1 / 18.21 / 1421 / Amino Acid Metabolism / 248 / 0.81**
histidine / 773 / 2 / 29.33 / 1909 / Amino Acid Metabolism / 154 / 0.58**
valine / 1182 / 1 / 12.43 / 1210 / Amino Acid Metabolism / 144 / 0.72**
phenylalanine / 994 / 1 / 23.02 / 1617 / Amino Acid Metabolism / 192 / 0.64**
tryptophan / 1148 / 1 / 34.85 / 2200 / Amino Acid Metabolism / 202 / 0.33**
asparagine / 236 / 1 / 24.07 / 1663 / Amino Acid Metabolism / 116 / 0.49*
tyrosine / 1153 / 1 / 29.75 / 1930 / Amino Acid Metabolism / 218 / 0.72*
proline / 145742 / 1 / 11.28 / 1170 / Amino Acid Metabolism / 70 / 0.52*
leucine / 857 / 1 / 14.03 / 1267 / Amino Acid Metabolism / 158 / 0.74*
lysine / 866 / 1 / 29.44 / 1914 / Amino Acid Metabolism / 174 / 0.77*
threonine / 205 / 1 / 17.13 / 1380 / Amino Acid Metabolism / 218 / 0.88*
aspartic acid / 424 / 1 / 20.58 / 1516 / Amino Acid Metabolism / 232 / 0.73*
4-guanidinobutyric acid / 500 / 1 / 20.76 / 1523 / Amino Acid Metabolism / 174 / 1.45*
isoleucine / 791 / 1 / 14.62 / 1288 / Amino Acid Metabolism / 158 / 0.81*
threitol / 169019 / 1 / 20.08 / 1495 / Carbohydrate Metabolism / 100 / 0.51**
lactulose / 16217605 / 2 / 38.18 / 2395 / Carbohydrate Metabolism / 361 / 0.69**
cellobiose / 17750876 / 1 / 42.75 / 2690 / Carbohydrate Metabolism / 204 / 0.54**
glucosaminic acid / 73563 / 1 / 31.69 / 2029 / Carbohydrate Metabolism / 291 / 0.68*
maltose / 10991489 / 1 / 43.21 / 2722 / Carbohydrate Metabolism / 204 / 0.79*
gluconic acid / 10690 / 2 / 29.24 / 1904 / Carbohydrate Metabolism / 275 / 1.25*
allantoin / 204 / 1 / 28.63 / 1874 / Carbohydrate Metabolism / 331 / 0.73*
ribofuranose-5-phosphate / 24794349 / 2 / 37.16 / 2334 / Carbohydrate Metabolism / 315 / 0.83*
myristic acid / 11005 / 1 / 28.02 / 1845 / Fatty acid Metabolism / 285 / 0.86*
pentadecanoic acid / 13849 / 1 / 30.02 / 1943 / Fatty acid Metabolism / 117 / 1.33*
glucose-6-phosphate / 439958 / 1 / 36.66 / 2305 / Glycolysis / 204 / 1.40*
3-indolelactic acid / 92904 / 2 / 34.62 / 2188 / Organic Acid Metabolism / 202 / 0.42**
pipecolic acid / 849 / 2 / 13.87 / 1261 / Organic Acid Metabolism / 84 / 2.50**
3-indoleacetic acid / 802 / 1 / 30.16 / 1950 / Organic Acid Metabolism / 202 / 1.22*
4,6-dioxoheptanoic acid / 5312 / 2 / 26.66 / 1780 / Organic Acid Metabolism / 257 / 0.82*
2,3,4-trihydroxybutyric acid / 439535 / 2 / 21.12 / 1538 / Organic Acid Metabolism / 292 / 1.36*
5-hydroxyindole-2-carboxylic acid / 88958 / 2 / 32.19 / 2055 / Organic Acid Metabolism / 378 / 1.22*
quinic acid / 6508 / 1 / 28.12 / 1849 / Phenol Metabolism / 345 / 1.57**
shikimic acid / 7057976 / 1 / 27.24 / 1807 / Phenol Metabolism / 204 / 1.26**
stigmasterol / 122544 / 2 / 50.34 / 3248 / Sterol and Vitamin Metabolism / 83 / 1.13**
succinic acid / 1110 / 1 / 15.26 / 1311 / TCA cycle / 147 / 1.56**
fumaric acid / 444972 / 1 / 16.27 / 1348 / TCA cycle / 245 / 1.45**
urea / 1176 / 2 / 23.35 / 1631 / Urea cycle / 403 / 0.61*
ornithine / 389 / 2 / 22.86 / 1610 / Urea cycle / 142 / 0.65**
citrulline / 9750 / 1 / 27.48 / 1819 / Urea cycle / 256 / 0.76*
dihydrosphingosine / 3126 / 2 / 38.75 / 2431 / / / 204 / 0.61**
porphine / 66868 / 2 / 15.59 / 1323 / / / 184 / 2.43*
a Identification level for metabolites according to Metabolomics Standards Initiative (MSI). 1: validation using reference standards; 2: identification using mass spectral library search. b The fold-change (8Km732/MH3301) is calculated by the comparison of average values between transgenic backcross breeding leaves (8Km732) and its non-transgenic parent (MH3301). *, ** and *** represent 0.01 < p < 0.05, 0.001 p < 0.01 and p < 0.001, respectively. p-value obtained based on nonparametric test (Mann−Whitney U test).
Table S2 Differential metabolites associated with pesticide stress in MH3301 leaf samples
Metabolite / PubChem CID / Identification Level a / Retention Time (min) / RI Index / Related pathways / Quantification Ions [m/z] / Fold-change b(MH3301 treated/MH3301control)
pre / 0.5d / 1d / 2d / 4d / 7d
serine / 5951 / 1 / 16.69 / 1357 / Amino Acid Metabolism / 204 / 1.4
aspartic acid / 424 / 1 / 18.38 / 1421 / Amino Acid Metabolism / 160 / 1.61 / 2.04
glutamic acid / 33032 / 1 / 23.22 / 1617 / Amino Acid Metabolism / 246 / 1.24
acetyl-L-serine / 25245438 / 2 / 14.50 / 1277 / Amino Acid Metabolism / 60 / 1.45
5-oxoproline / 7405 / 2 / 20.77 / 1515 / Amino Acid Metabolism / 156 / 1.23
isoleucine / 791 / 1 / 14.84 / 1289 / Amino Acid Metabolism / 158 / 0.7
4-guanidinobutyric acid / 500 / 1 / 21.01 / 1525 / Amino Acid Metabolism / 174 / 0.76
glycine / 750 / 1 / 15.15 / 1300 / Amino Acid Metabolism / 174 / 0.87
methoxytryptamine / 1833 / 2 / 39.62 / 2471 / Amino Acid Metabolism / 174 / 0.61
dehydroascorbic acid / 440667 / 2 / 28.61 / 1863 / / / 316 / 1.31
ascorbic acid / 54670067 / 1 / 30.14 / 1939 / / / 332 / 1.11
mucic acid / 607 / 2 / 31.96 / 2032 / Carbohydrate Metabolism / 333 / 1.4 / 1.63 / 1.43
inositol-3-phosphate / 440194 / 2 / 38.47 / 2399 / Carbohydrate Metabolism / 318 / 1.28
erythritol / 222285 / 1 / 20.49 / 1504 / Carbohydrate Metabolism / 217 / 1.47
glucoheptonic acid / 101713 / 2 / 31.14 / 1985 / Carbohydrate Metabolism / 103 / 0.45 / 0.4 / 0.68
galactinol / 44629828 / 2 / 48.84 / 3114 / Carbohydrate Metabolism / 204 / 1.26 / 0.43 / 0.54
fructose / 5984 / 1 / 28.85 / 1875 / Carbohydrate Metabolism / 307 / 0.83 / 0.77
cellobiose / 17750876 / 1 / 42.99 / 2691 / Carbohydrate Metabolism / 204 / 0.75 / 0.75
arabinose / 854 / 1 / 24.15 / 1658 / Carbohydrate Metabolism / 103 / 1.37 / 1.26
sorbitol / 5780 / 1 / 29.89 / 1926 / Carbohydrate Metabolism / 319 / 1.19 / 1.25
lactobionic acid / 7314 / 2 / 36.08 / 2259 / Carbohydrate Metabolism / 204 / 0.8
lactulose / 16217605 / 2 / 35.10 / 2202 / Carbohydrate Metabolism / 217 / 1.32
trehalose-6-phosphate / 122336 / 2 / 47.56 / 2999 / Carbohydrate Metabolism / 195 / 0.84
arbutin / 440936 / 2 / 41.26 / 2576 / Carbohydrate Metabolism / 254 / 0.89
xylose / 644160 / 1 / 23.99 / 1651 / Carbohydrate Metabolism / 307 / 1.18
tagatose / 92092 / 1 / 28.32 / 1842 / Carbohydrate Metabolism / 307 / 1.67
mannitol / 6251 / 1 / 29.76 / 1920 / Carbohydrate Metabolism / 319 / 0.69
maltose / 10991489 / 1 / 43.45 / 2722 / Carbohydrate Metabolism / 204 / 0.83 / 0.67
threitol / 169019 / 1 / 20.31 / 1496 / Carbohydrate Metabolism / 217 / 0.85
ribose / 90428 / 1 / 24.48 / 1672 / Carbohydrate Metabolism / 103 / 0.86
galactose / 6036 / 1 / 29.01 / 1882 / Carbohydrate Metabolism / 319 / 1.25 / 1.34 / 0.65 / 0.79
3-phosphoglycerate / 724 / 2 / 27.33 / 1801 / Glycolysis / 357 / 1.74 / 2.52
glucose-6-phosphate / 439958 / 1 / 36.90 / 2306 / Glycolysis / 204 / 0.71 / 0.68 / 0.72
glucose / 5793 / 1 / 29.51 / 1907 / Glycolysis / 319 / 0.81 / 0.79
oxalic acid / 971 / 1 / 10.35 / 1132 / Organic Acid Metabolism / 147 / 1.55
phosphoric acid / 1004 / 1 / 14.28 / 1269 / Organic Acid Metabolism / 299 / 2.05
malonic acid / 867 / 1 / 12.37 / 1201 / Organic Acid Metabolism / 147 / 0.78
glycerol-1-phosphate / 754 / 1 / 26.37 / 1757 / / / 357 / 1.22
glyceric acid / 752 / 1 / 15.88 / 1327 / Organic Acid Metabolism / 292 / 0.74
3-hydroxypropanoic acid / 68152 / 2 / 10.64 / 1142 / Organic Acid Metabolism / 147 / 0.71
sphingosine / 5280335 / 2 / 36.66 / 2292 / Amine Metabolism / 204 / 0.65
salicylic acid / 338 / 1 / 20.47 / 1502 / Phenol Metabolism / 267 / 0.85 / 0.85
benzoic acid, 2,6-dihydroxy / 9338 / 2 / 26.24 / 1751 / Phenol Metabolism / 355 / 1.14
tocopherol / 14985 / 2 / 48.97 / 3123 / Phenol Metabolism / 237 / 0.77
resorcinol / 5054 / 2 / 17.78 / 1397 / Phenol Metabolism / 239 / 0.8
2-(4-hydroxyphenyl)ethanol / 10393 / 2 / 22.03 / 1567 / Phenol Metabolism / 179 / 0.77
gentisic acid / 3469 / 2 / 26.64 / 1769 / Phenol Metabolism / 355 / 0.71
shikimic acid / 7057976 / 1 / 27.47 / 1809 / Phenol Metabolism / 204 / 2.39 / 1.73
quinic acid / 6508 / 1 / 28.35 / 1851 / Phenol Metabolism / 345 / 1.8
stigmasterol / 122544 / 2 / 50.64 / 3248 / Sterol and Vitamin Metabolism / 83 / 1.14
campesterol / 312822 / 2 / 50.33 / 3225 / Sterol and Vitamin Metabolism / 382 / 1.12 / 1.12
sitosterol / 222284 / 2 / 51.40 / 3304 / Sterol and Vitamin Metabolism / 396 / 1.19 / 1.1
nicotinamide / 936 / 2 / 19.76 / 1475 / Sterol and Vitamin Metabolism / 179 / 1.26 / 0.82
phytol / 5366244 / 2 / 34.52 / 2170 / Terpene Metabolism / 143 / 0.82 / 0.79 / 0.83
isocitric acid / 1198 / 1 / 27.66 / 1817 / TCA Cycle / 245 / 1.48
citric acid / 311 / 1 / 27.58 / 1814 / TCA Cycle / 273 / 0.81
aconitic acid / 643757 / 1 / 26.15 / 1747 / TCA Cycle / 229 / 0.78
a Identification level for metabolites according to Metabolomics Standards Initiative (MSI). 1: validation using reference standards; 2: identification using mass spectral library search. b The fold-change was obtained by the comparison of average values between treated and untreated MH3301 leaves. Red and blue represent the level of metabolite in treated groups was higher and lower than that in untreated samples, respectively.
Table S3 Differential metabolites associated with pesticide stress in 8Km732 leaf samples
Name / PubChem CID / Identification Level a / Retention Time (min) / RI Index / Related pathways / Quantification Ions [m/z] / Fold-change b(8Km732 treated/8Km732 control)
pre / 0.5d / 1d / 2d / 4d / 7d
stearic acid / 5281 / 1 / 35.78 / 2241 / Fatty Acid Metabolism / 117 / 1.22
palmitic acid / 985 / 1 / 32.19 / 2044 / Fatty Acid Metabolism / 117 / 1.22
proline / 145742 / 1 / 11.52 / 1172 / Amino Acid Metabolism / 70 / 1.22
serine / 5951 / 1 / 16.69 / 1357 / Amino Acid Metabolism / 204 / 1.62 / 1.65
glutamic acid / 33032 / 1 / 23.22 / 1617 / Amino Acid Metabolism / 246 / 1.28
acetyl-L-serine / 25245438 / 2 / 14.50 / 1277 / Amino Acid Metabolism / 60 / 1.5
5-oxoproline / 7405 / 2 / 20.77 / 1515 / Amino Acid Metabolism / 156 / 1.19 / 1.2 / 1.3
ethanolamine / 700 / 1 / 14.12 / 1263 / Amino Acid Metabolism / 84 / 1.65 / 3 / 1.7
glycine / 750 / 1 / 15.15 / 1300 / Amino Acid Metabolism / 174 / 1.3
N-methylglutamic acid / 439377 / 2 / 14.59 / 1280 / Amino Acid Metabolism / 98 / 1
valine / 1182 / 1 / 12.65 / 1212 / Amino Acid Metabolism / 144 / 0.7
threonine / 205 / 1 / 17.35 / 1382 / Amino Acid Metabolism / 218 / 1.45
methoxytryptamine / 1833 / 2 / 39.62 / 2471 / Amino Acid Metabolism / 174 / 1.09
5-hydroxytryptamine / 5202 / 2 / 18.46 / 1424 / Amino Acid Metabolism / 174 / 1.09
alanine / 5950 / 1 / 9.42 / 1100 / Amino Acid Metabolism / 116 / 1.29 / 1.17
dehydroascorbic acid / 440667 / 2 / 28.61 / 1863 / / / 316 / 0.8
ascorbic acid / 54670067 / 1 / 30.14 / 1939 / / / 332 / 0.9
melezitose / 92817 / 2 / 53.59 / 3468 / Carbohydrate Metabolism / 361 / 1.16
inositol-3-phosphate / 440194 / 2 / 38.47 / 2399 / Carbohydrate Metabolism / 318 / 1.3
fructose 6-phosphate / 69507 / 2 / 36.73 / 2296 / Carbohydrate Metabolism / 315 / 1.4
mannose / 18950 / 1 / 28.92 / 1878 / Carbohydrate Metabolism / 95 / 1.34 / 1.3
lactulose / 16217605 / 2 / 35.10 / 2202 / Carbohydrate Metabolism / 217 / 1.3
galactinol / 44629828 / 2 / 48.84 / 3114 / Carbohydrate Metabolism / 204 / 0.55 / 0.7
galactose / 6036 / 1 / 29.01 / 1882 / Carbohydrate Metabolism / 319 / 0.8
fructose / 5984 / 1 / 28.85 / 1875 / Carbohydrate Metabolism / 307 / 0.83 / 0.71 / 0.8
ribose / 90428 / 1 / 24.48 / 1672 / Carbohydrate Metabolism / 103 / 1.2
rhamnose / 439548 / 1 / 25.32 / 1710 / Carbohydrate Metabolism / 117 / 1.3
arabinose / 854 / 1 / 24.15 / 1658 / Carbohydrate Metabolism / 103 / 1.2
cellobiose / 17750876 / 1 / 42.99 / 2691 / Carbohydrate Metabolism / 204 / 0.71 / 0.7 / 0.67 / 0.8
glucoheptonic acid / 101713 / 2 / 31.14 / 1985 / Carbohydrate Metabolism / 103 / 0.57 / 0.5 / 0.58
3-phosphoglycerate / 724 / 2 / 27.33 / 1801 / Glycolysis / 357 / 1.8
glucose / 5793 / 1 / 29.51 / 1907 / Glycolysis / 319 / 0.83 / 0.69 / 0.8
glucose-6-phosphate / 439958 / 1 / 36.90 / 2306 / Glycolysis / 204 / 0.68 / 0.7 / 0.81 / 1.4
glyceric acid / 752 / 1 / 15.88 / 1327 / Organic Acid Metabolism / 292 / 1.49 / 0.8
glycolic acid / 757 / 1 / 8.70 / 1074 / Organic Acid Metabolism / 147 / 1.4
lactobionic acid / 7314 / 2 / 36.08 / 2259 / Organic Acid Metabolism / 204 / 0.8 / 0.8
gluconic acid / 10690 / 2 / 31.05 / 1985 / Organic Acid Metabolism / 217 / 0.9 / 0.7
acetol / 8299 / 2 / 21.53 / 1546 / / / 219 / 0.9
3-hydroxypropanoic acid / 68152 / 2 / 10.64 / 1142 / Organic Acid Metabolism / 147 / 0.8
phytosphingosine / 122121 / 2 / 34.15 / 2149 / / / 204 / 0.9
2-hydroxypyridine / 8871 / 1 / 7.55 / 1032 / / / 152 / 0.8
isocitric lactone / 98259 / 2 / 25.19 / 1703 / / / 157 / 0.8 / 0.7
2,3-butanediol / 262 / 2 / 7.65 / 1036 / / / 117 / 1.94
propylene glycol / 1030 / 2 / 6.56 / 997 / / / 117 / 0.87 / 1.25
4-hydroxypyridine / 12290 / 1 / 10.86 / 1150 / / / 152 / 0.78
gentisic acid / 3469 / 2 / 26.64 / 1769 / Phenol Metabolism / 355 / 0.61 / 0.6
2-(4-hydroxyphenyl)ethanol / 10393 / 2 / 22.03 / 1567 / Phenol Metabolism / 179 / 0.9 / 0.8
benzoic acid, 2,6-dihydroxy / 9338 / 2 / 26.24 / 1751 / Phenol Metabolism / 355 / 0.79
2,2-bipyridine / 1474 / 2 / 18.92 / 1442 / Phenol Metabolism / 156 / 0.64
shikimic acid / 7057976 / 1 / 27.47 / 1809 / Shikimate Pathway and Phenylpropanoids / 204 / 1.21