succinic acid / -0.0794 / 0.1071 / 0.0798 / 0.2232 / 0.0477 / -0.0376
galactinol / -0.0677 / -0.1633 / 0.1979 / -0.0169 / -0.0565 / 0.0744
N-acetylglutamate / -0.0367 / 0.0742 / 0.1105 / 0.1669 / -0.2436 / 0.0314
inositol-1-P / -0.0294 / -0.0753 / 0.2338 / 0.1638 / 0.2105 / 0.0735
2-methyl-malic acid / -0.0293 / 0.2078 / -0.1303 / 0.0909 / -0.0447 / -0.1369
galactitol (put) / -0.0235 / 0.0516 / 0.1906 / 0.2227 / 0.0693 / 0.1247
maleic acid / -0.0145 / 0.2426 / -0.1348 / 0.1261 / -0.0568 / -0.0586
2,3-dihydrobutanedioic acid / -0.0119 / 0.1395 / -0.0602 / 0.2095 / 0.0684 / 0.0872
beta-alanine / 0.0097 / 0.2439 / -0.1523 / 0.0441 / 0.1260 / 0.1007
glycerol-3-P / 0.0102 / 0.1945 / -0.0919 / 0.1686 / -0.0368 / 0.1113
monomethylphosphate / 0.0113 / 0.1055 / 0.0785 / 0.1917 / -0.3232 / -0.0090
3PGA / 0.0214 / 0.1410 / -0.0831 / 0.1605 / 0.0244 / 0.0320
digalactosylglycerol / 0.0245 / -0.0048 / 0.2059 / 0.0987 / 0.0641 / 0.1476
t-ferulic_acid / 0.0246 / -0.1348 / 0.1703 / -0.0020 / -0.0383 / -0.0890
GABA / 0.0269 / 0.0725 / 0.0017 / -0.0048 / 0.1166 / -0.1493
tetracosanoic acid / 0.0270 / 0.0303 / 0.0958 / 0.1130 / 0.2375 / 0.1534
serine / 0.0357 / 0.1913 / -0.2333 / -0.0621 / 0.0047 / 0.0936
frc-6-P / 0.0578 / 0.1272 / 0.0137 / 0.0902 / 0.2438 / 0.0607
threonic acid-1,4-lactone / 0.0584 / 0.1201 / -0.0088 / 0.0402 / 0.0591 / -0.1453
threonic acid / 0.0623 / 0.1444 / 0.0870 / 0.1061 / 0.1311 / -0.0796
aspartate / 0.0629 / 0.0959 / -0.2347 / -0.0765 / -0.0761 / 0.0175
proline / 0.0645 / 0.0256 / 0.0682 / -0.0977 / -0.0266 / 0.0434
1-methyl-beta-D-galactose(put) / 0.0648 / 0.0710 / 0.0568 / -0.0737 / -0.0412 / 0.1115
glc-6-P / 0.0683 / 0.0865 / 0.0890 / 0.0737 / 0.2272 / 0.0762
hexacosanol / 0.0708 / -0.0304 / -0.0469 / -0.1159 / 0.2491 / 0.0474
benzoic acid / 0.0722 / 0.0259 / 0.0852 / -0.0403 / -0.0965 / -0.2649
1,6-anhydroglucose / 0.0729 / 0.1295 / 0.1357 / 0.0362 / -0.2165 / -0.0039
phosphate / 0.0735 / -0.0066 / 0.0476 / 0.1047 / -0.1136 / 0.2513
putrescine / 0.0736 / 0.1502 / -0.1786 / -0.0574 / -0.1393 / 0.0448
UL028 / 0.0748 / 0.0008 / -0.0019 / -0.1624 / 0.0870 / -0.0071
glyceric acid / 0.0752 / 0.1601 / 0.0224 / 0.0962 / -0.0919 / -0.1196
isocitrate / 0.0756 / -0.0347 / -0.0021 / -0.0363 / -0.0749 / 0.1990
xylitol / 0.0758 / 0.0920 / -0.0609 / -0.0557 / 0.2414 / 0.0802
9(Z)-18:1 / 0.0769 / 0.0708 / 0.1751 / -0.0732 / -0.1433 / -0.2486
9,12(Z,Z)-18:2 / 0.0777 / 0.0158 / 0.0775 / 0.0060 / 0.1680 / 0.0713
fumaric_acid / 0.0780 / 0.1375 / 0.1306 / 0.0339 / -0.0784 / -0.1280
threonine / 0.0793 / 0.2160 / -0.0022 / 0.1133 / -0.0585 / 0.0556
raffinose / 0.0803 / -0.0284 / 0.1708 / 0.0358 / -0.0704 / 0.2180
fructose / 0.0804 / 0.0867 / 0.0965 / -0.0412 / -0.0775 / -0.2138
uracil / 0.0821 / -0.1339 / 0.0813 / -0.0616 / -0.0055 / -0.0033
methionine / 0.0826 / -0.0521 / -0.0611 / 0.1592 / -0.0450 / -0.0154
aminoisobutyric acid / 0.0837 / -0.0766 / -0.0781 / 0.0887 / 0.1398 / -0.1795
salicylic acid / 0.0869 / -0.0036 / 0.1491 / -0.0402 / 0.0322 / -0.0686
tryptophan / 0.0881 / -0.1007 / -0.0593 / 0.1287 / 0.0665 / -0.0765
histidine / 0.0885 / -0.0229 / -0.0045 / -0.0691 / -0.0376 / 0.1619
decanedioic acid / 0.0889 / -0.1149 / -0.0832 / 0.1208 / -0.0333 / 0.0471
docosanoic acid / 0.0905 / -0.1439 / -0.0613 / 0.1251 / -0.0090 / -0.0263
glycine / 0.0909 / -0.1216 / -0.0816 / 0.1913 / 0.0148 / -0.1348
Metabolites / LS 1 / LS 2 / LS 3 / LS 4 / LS 5 / LS 6
unknown_071306a / 0.0909 / -0.0477 / -0.0234 / 0.0965 / -0.0842 / 0.2078
tyramine / 0.0911 / 0.0637 / 0.0637 / -0.0838 / 0.0368 / -0.0692
pentadecanoic acid / 0.0928 / 0.0741 / 0.1062 / 0.0260 / 0.0318 / -0.1006
2-oxobutryic acid / 0.0954 / -0.0157 / -0.0143 / -0.1845 / -0.0274 / -0.0552
erythronic acid (put) / 0.0961 / 0.1338 / 0.1170 / 0.1293 / -0.0948 / -0.0529
20:0 / 0.0967 / 0.0111 / 0.1663 / -0.0118 / -0.0873 / 0.0115
asparagine / 0.0984 / -0.1174 / -0.0605 / 0.0985 / -0.0136 / 0.0614
melezitose / 0.0987 / 0.0437 / -0.1064 / -0.1160 / -0.1554 / 0.0058
isoleucine / 0.0992 / -0.0910 / -0.0882 / 0.1799 / 0.0183 / -0.0763
glutamine / 0.0996 / -0.0577 / -0.0312 / 0.0033 / -0.0464 / 0.1381
muconic acid / 0.1000 / -0.1582 / -0.0666 / 0.1173 / -0.0250 / 0.0086
phytanic acid / 0.1000 / -0.0318 / -0.0446 / 0.0460 / -0.0160 / -0.0016
1-kestose / 0.1001 / 0.0718 / -0.0989 / -0.1214 / -0.1170 / 0.0289
malic_acid / 0.1006 / -0.0033 / 0.1478 / 0.0449 / -0.1070 / 0.1504
alanine / 0.1013 / -0.1077 / -0.0924 / 0.1756 / 0.0170 / -0.1093
octadecanol / 0.1020 / 0.0458 / 0.1610 / -0.0526 / 0.0322 / -0.1423
1-monohexadecanoglycerol / 0.1021 / 0.0382 / 0.1907 / 0.0262 / -0.0273 / -0.0377
6-kestose / 0.1050 / 0.0464 / 0.0722 / 0.0194 / -0.0129 / 0.1258
acetohydroxamic acid / 0.1065 / -0.0339 / -0.0488 / -0.0833 / -0.1222 / 0.1710
oxalic acid / 0.1096 / -0.0835 / -0.0429 / 0.0665 / -0.0331 / 0.0670
glycerol-2-P / 0.1098 / 0.0198 / -0.0473 / -0.1377 / 0.1088 / -0.0174
valine / 0.1105 / -0.0989 / -0.0935 / 0.1637 / 0.0015 / -0.0442
urea / 0.1106 / -0.1071 / 0.0112 / 0.0432 / -0.0481 / -0.0245
glycerol_ / 0.1119 / -0.0777 / -0.0489 / 0.0468 / 0.0276 / -0.1315
shikimic acid / 0.1124 / 0.1275 / 0.0083 / -0.0175 / 0.0859 / -0.0386
5-oxoproline / 0.1124 / -0.0541 / -0.0575 / -0.0243 / -0.0422 / 0.0810
U_021706b_24.04 / 0.1127 / 0.0169 / 0.0717 / -0.0176 / 0.0253 / -0.0120
itaconic acid / 0.1148 / -0.0679 / -0.0672 / -0.0580 / -0.0068 / -0.0516
leucine / 0.1148 / -0.0614 / -0.0957 / 0.1064 / 0.0127 / -0.0758
lysine / 0.1150 / -0.0795 / -0.0693 / 0.1053 / 0.0181 / -0.0876
9,12,15(Z,Z,Z)-18:3 / 0.1151 / 0.0557 / 0.0821 / 0.0200 / 0.0023 / 0.0482
U_021706b_40.484 / 0.1153 / -0.0377 / 0.1114 / -0.0502 / 0.0333 / -0.0349
quinic_acid / 0.1156 / 0.1279 / -0.0040 / -0.0001 / 0.0226 / 0.0460
glucaric acid / 0.1158 / 0.0177 / 0.0035 / -0.0076 / -0.1317 / -0.0254
ribose / 0.1159 / -0.0416 / 0.0398 / 0.0235 / 0.0659 / -0.0979
tyrosine / 0.1161 / -0.0642 / -0.0495 / 0.0228 / 0.0336 / 0.0144
phenylalanine / 0.1172 / -0.0797 / -0.0739 / 0.1348 / 0.0324 / -0.0748
U_021706b_33.7 / 0.1181 / 0.1208 / -0.0250 / -0.0246 / -0.0245 / 0.0121
homoserine / 0.1183 / -0.0775 / -0.0700 / 0.1387 / 0.0237 / -0.0358
guanine / 0.1186 / -0.0554 / -0.0453 / -0.0541 / -0.0634 / 0.0177
citric acid / 0.1190 / 0.1001 / 0.0033 / -0.0125 / 0.0595 / 0.0245
lactic acid / 0.1190 / -0.0428 / -0.0295 / -0.0160 / -0.0786 / 0.1392
U_042806a_27.94_sugar / 0.1202 / 0.0927 / -0.0301 / -0.0575 / 0.0925 / 0.0031
xylose / 0.1207 / 0.0781 / 0.0082 / -0.0921 / 0.0792 / 0.0529
malonic acid / 0.1225 / -0.0758 / -0.0352 / 0.0214 / -0.0277 / 0.0626
oxaloacetic acid / 0.1231 / -0.0616 / -0.0721 / 0.0131 / -0.0090 / -0.0191
4-hydroxycinnamic acid / 0.1235 / -0.0370 / -0.0350 / -0.0683 / -0.0321 / 0.0721
glycolic acid / 0.1235 / -0.0792 / -0.0434 / -0.0367 / -0.0219 / -0.0458
glutamate / 0.1239 / -0.0993 / -0.0512 / 0.0810 / 0.0017 / -0.0146
Metabolites / LS 1 / LS 2 / LS 3 / LS 4 / LS 5 / LS 6
U_021706b_23.87_sugar_acid / 0.1242 / 0.0595 / -0.0227 / -0.0489 / 0.0143 / 0.0208
18:00 / 0.1242 / -0.0009 / 0.1295 / -0.0323 / -0.0140 / 0.0093
arabinose / 0.1242 / 0.0590 / 0.0317 / -0.0645 / 0.1196 / 0.0184
tetradecanoic acid / 0.1243 / -0.0694 / -0.0387 / -0.0150 / -0.0263 / 0.0743
16:0 / 0.1249 / 0.0323 / 0.0900 / -0.0669 / 0.0739 / -0.0394
galacturonic acid (put) / 0.1253 / 0.0559 / 0.0360 / -0.0594 / 0.0066 / 0.0037
sucrose / 0.1254 / 0.0233 / 0.0630 / -0.0738 / 0.0329 / 0.0246
glucose / 0.1261 / 0.0926 / 0.0015 / -0.0537 / 0.0227 / 0.0087
aconitic acid / 0.1265 / 0.0135 / 0.0223 / -0.0502 / -0.0231 / 0.0055
1-monooctodecanoglycerol / 0.1268 / -0.0094 / 0.0992 / -0.0508 / 0.0651 / -0.0176
myo-inositol / 0.1276 / 0.0266 / 0.0058 / -0.0993 / -0.0175 / 0.0064
heptadecanoic acid / 0.1280 / -0.0775 / -0.0417 / -0.0204 / 0.0145 / -0.0512
Supplemental Table 2 Statistical significance of the Discriminant Functions
Function / Wilks' Lambda (L) / Chi-square / df / p-levelRemoved
0 / 0.000000 / 565.188500 / 168.000000 / 0.000000
1 / 0.000036 / 296.853400 / 110.000000 / 0.000000
2 / 0.037466 / 95.245000 / 54.000000 / 0.000455
To test for the statistical significance of the discriminant function, the Wilks' test (as measured by the Wilks' lambda "L" was used. It is based on the discrimination that remains in the system before deriving that function. If "L" approaches 1.0, there is no discrimination. If "L" approaches 0, there is maximum discrimination. The statistical significance of "L"is tested by converting it into an approximation of the Chi-square distribution and the probability calculated. The table shows that the separation of genotypes for the first and second discriminant functions were maximal and there was almost chance that the separation arose from chance alone.
Supplemental Table 3. Compounds with altered correlative patterns when compared between genotypes (r<0.90)
Dx5-G BAR-C / Dx5-GBAR-D / Dx5-J
BAR-C / Dx5-J
BAR-D / Dx5-G
Dx5-J / BAR-C
BAR-D
1,6-anhydroglucose / O / O / O
1-methyl-beta-D-galactose (put) / O / O / O / O / O
1-monohexadecanoglycerol / / /
2-methyl-malic acid / O / O
2,3-dihydroxybutanedioic acid / / /
3PGA / O / O / O / O
6-kestose / /
9,12(Z,Z)-18:1 / O / O
9,12(Z,Z)-18:2 / O / O / O / O
aminoisobutyric acid / / /
aspartate / O / O / O / O / O
benzoic acid / O / O / O / O
Beta-alanine / O / O / O
digalactosylglycerol / O / O / O / O
20:0 / / /
erythronic acid (put) / / / /
Fructose-6-P / O / O / O
fumaric_acid / O / O / O
GABA / / / /
galactitol / O / O
Glc-6-P / O / O / O / O
glyceric acid / / / / /
Glycerol-3-P / / / /
Inositol-1-P / O / O / O / O
isocitrate / O / O / O
isoleucine /
malic_acid / /
methionine / O / O / O / O
monomethylphosphate / O / O / O / O / O
N-acetylglutamine / / / /
oxalic acid / /
pentadecanoic acid / / /
phosphate / O / O / O / O
Phytanic acid / /
proline / / / / /
putrescine / /
raffinose /
serine / O / O / O
shikimic acid / / / /
tetracosanoic acid* / / / / /
t-ferulic_acid / O / O / O / O
threonic acid / / / /
threonic acid-1,4-lactone / / / /
threonine / O / O
UL028 /
Unknown_071306a / / /
xylitol /
Correlations were calculated using Pearson rank correlation coefficients. Of the 47 metabolites with altered profiles, 22 varied when BAR-C and BAR-D were compared to each other and may have arisen from the transformation process. Twenty-six metabolites showed altered correlations when the Dx5-transformed genotypes were aligned to each other or to any of the bar-only transformants (controls). Key is as follows, metabolites with an altered profile between: (i) the two bar-only transformants compared to each other, and, compared to each Dx5-transformed genotypes "O"; (ii) the Dx5-transformed genotypes against each other and against each control ; (iii) both Dx5-transformed genotypes against each control only ; (iv) either of the two Dx5-transformed genotypes against either control ; (v) Dx5-G compared with either control ; (vi) Dx5-J and either control . Three compounds, UL028, raffinose, and isoleucine, only changed in one comparison of aDx5-transformed genotype with a bar-only line but were included
Supplemental Table 4. Compounds significantly different (t-test; P<0.05 in black or p<0.0005 in red) between the two bar-only transformants at 7, 14 and 21DPA
7 DPAav. ratio
Compounds / Va/Vb 7d / t-test
4-hydroxycinnamic acid / 1.573 / 0.01065
acetohydroxamic acid / 3.331 / 0.00009
aconitic acid / 1.088 / 0.61952
aspartate / 0.645 / 0.01755
beta-alanine / 0.597 / 0.02630
frc-6-P / 0.753 / 0.03191
galactinol / 1.590 / 0.03988
glc-6-P / 0.760 / 0.00850
guanine / 1.660 / 0.00673
lactic acid / 1.843 / 0.00247
unknown_071306a / 2.041 / 0.00237
14 DPA
av. ratio
Compounds / Va/Vb 14d / t-test
1-kestose / 2.011 / 0.00479
acetohydroxamic acid / 1.882 / 0.02213
aspartate / 1.539 / 0.01747
glucaric acid / 1.473 / 0.02157
lactic acid / 1.673 / 0.01681
melezitose / 1.979 / 0.00475
21 DPA
av. ratio
Compounds / Va/Vb 21d / t-test
1,6-anhydroglucose / 1.968 / 0.00046
1-kestose / 2.945 / 0.03969
1-methyl-beta-D-galactose(put) / 2.061 / 0.01222
3PGA / 1.935 / 0.00988
4-hydroxycinnamic acid / 1.547 / 0.00122
acetohydroxamic acid / 2.362 / 0.00054
aspartate / 2.149 / 0.00572
glc-6-P / 0.533 / 0.01272
glycerol-3-P / 1.758 / 0.00302
guanine / 1.678 / 0.01574
histidine / 2.336 / 0.03976
lactic acid / 1.712 / 0.00002
maleic acid / 2.143 / 0.00150
melezitose / 3.204 / 0.03165
putrescine / 5.026 / 0.00312
serine / 2.014 / 0.04555
t-ferulic_acid / 0.607 / 0.02479
U_021706b_33.7 / 2.318 / 0.00017
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