Table 1.
m\z | RI | PC1 (49%) loadings | PC2 (27%) loadings | N. Ph. average response | 24h darkness average response | 72h darkness average response | 24h/N. Ph. response ratio | 72h/N. Ph. response ratio | P-value | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Sugars | ||||||||||||||||||||
Sucrose | 361 | 2385 | –0.5010 | 0.3580 | 8.679 | 7.866 | 3.276 | 0.91 a | 0.38 b | <0.001 | ||||||||||
Raffinose | 87 | 3056 | –0.0197 | –0.0103 | 0.268 | 0.060 | 0.026 | 0.22 | 0.10 | 0.010 | ||||||||||
Maltose | 361 | 2495 | 0.0005 | –0.0023 | 0.059 | 0.051 | 0.068 | 0.86 | 1.14 | 0.833 | ||||||||||
Trehalose | 361 | 2505 | 0.0144 | –0.0178 | 0.329 | 0.413 | 0.492 | 1.25 | 1.49 | 0.310 | ||||||||||
d-Melezitose | 361 | 3153 | –0.0380 | –0.0010 | 0.558 | 0.231 | 0.069 | 0.41 a | 0.12 b | <0.001 | ||||||||||
Melibiose | 361 | 2512 | –0.0046 | 0.0043 | 0.216 | 0.146 | 0.155 | 0.67 | 0.72 | 0.208 | ||||||||||
Cellobiose | 361 | 2252 | –0.0002 | –0.0006 | 0.020 | 0.016 | 0.015 | 0.79 | 0.77 | 0.870 | ||||||||||
Maltotriose | 361 | 3034 | –0.0031 | 0.0008 | 0.039 | 0.015 | 0.002 | 0.40 | 0.06 | 0.012 | ||||||||||
d-Lyxose | 217 | 1389 | –0.0012 | 0.0001 | 0.024 | 0.019 | 0.011 | 0.80 | 0.48 | 0.393 | ||||||||||
d-Glucose | 319 | 1614 | –0.0187 | 0.0033 | 0.237 | 0.088 | 0.000 | 0.37 a | 0.00 b | 0.047 | ||||||||||
Amino acids | ||||||||||||||||||||
l-Cysteine | 220 | 1250 | 0.0047 | –0.0030 | 0.000 | 0.042 | 0.047 | Dark sp. | Dark sp. | 0.529 | ||||||||||
l-Ornithine | 142 | 1310 | 0.0150 | –0.0091 | 0.000 | 0.000 | 0.185 | ND | Dark sp. | – | ||||||||||
l-Homoserine | 218 | 1145 | 0.0103 | 0.0018 | 0.017 | 0.062 | 0.129 | 3.70 | 7.74 | 0.028 | ||||||||||
l-Proline | 142 | 983 | –0.0048 | 0.0025 | 0.106 | 0.074 | 0.042 | 0.69 | 0.40 | 0.603 | ||||||||||
l-Asparagine | 231 | 1376 | 0.4410 | 0.1160 | 11.782 | 14.955 | 17.094 | 1.27 a | 1.45 b | <0.001 | ||||||||||
β-Alanine | 248 | 1116 | 0.0057 | –0.0005 | 0.052 | 0.060 | 0.119 | 1.16 a | 2.30 b | 0.009 | ||||||||||
l-Glutamic acid | 246 | 1319 | 0.1840 | 0.8760 | 2.066 | 5.274 | 4.310 | 1.28 | 1.15 | 0.211 | ||||||||||
β-Cyano-l-alanine | 141 | 1064 | 0.0022 | 0.0020 | 0.190 | 0.201 | 0.187 | 1.06 | 0.98 | 0.959 | ||||||||||
l-Alanine | 116 | 798 | –0.0252 | 0.0160 | 0.400 | 0.397 | 0.119 | 0.99 a | 0.30 b | 0.004 | ||||||||||
l-Serine | 204 | 1055 | 0.0262 | –0.0019 | 0.330 | 0.430 | 0.673 | 1.31 a | 2.04 b | 0.018 | ||||||||||
l-Aspartic acid | 232 | 1216 | 0.3470 | –0.0640 | 0.873 | 2.162 | 4.931 | 2.48 a | 5.65 b | <0.001 | ||||||||||
l-Lysine | 317 | 1618 | 0.0722 | –0.0294 | 0.212 | 0.334 | 1.066 | 1.57 a | 5.02 b | <0.001 | ||||||||||
l-Tyrosine | 218 | 1635 | 0.0674 | 0.0217 | 0.000 | 0.682 | 0.778 | Dark sp. | Dark sp. | 0.059 | ||||||||||
l-Methionine | 176 | 1208 | 0.0066 | –0.0036 | 0.000 | 0.000 | 0.070 | ND. | Dark sp. | – | ||||||||||
l-Histidine | 154 | 1618 | 0.1240 | –0.0340 | 0.065 | 0.186 | 1.450 | 2.88 a | 22.45 b | <0.001 | ||||||||||
l-Valine | 144 | 903 | 0.0888 | –0.0378 | 0.106 | 0.280 | 1.185 | 2.65 a | 11.21 b | <0.001 | ||||||||||
l-Threonine | 218 | 1083 | 0.0211 | –0.0002 | 0.166 | 0.343 | 0.434 | 2.07 | 2.62 | <0.001 | ||||||||||
l-Tryptophan | 202 | 1918 | 0.0878 | –0.0262 | 0.046 | 0.350 | 1.120 | 7.64 a | 24.41 b | <0.001 | ||||||||||
dl-Isoleucine | 158 | 982 | 0.0768 | 0.0058 | 0.034 | 0.297 | 0.875 | 8.70 a | 25.58 b | <0.001 | ||||||||||
l-Glutamine | 156 | 1470 | –0.0184 | –0.0036 | 0.411 | 0.129 | 0.130 | 0.31 | 0.32 | <0.001 | ||||||||||
Glycine | 174 | 993 | 0.0103 | 0.0021 | 0.027 | 0.097 | 0.146 | 3.55 | 5.35 | 0.028 | ||||||||||
Organic acids | ||||||||||||||||||||
Dehydroascorbic acid | 173 | 1553 | –0.0025 | –0.0019 | 0.036 | 0.000 | 0.000 | 0.00 | 0.00 | – | ||||||||||
Shikimic acid | 204 | 1517 | –0.0019 | 0.0003 | 0.032 | 0.000 | 0.000 | 0.00 | 0.00 | – | ||||||||||
6-Hydroxycaproic acid | 147 | 1010 | 0.0017 | –0.0011 | 0.018 | 0.031 | 0.033 | 1.68 | 1.81 | 0.626 | ||||||||||
l-Ascorbic acid | 332 | 1659 | –0.0107 | –0.0086 | 0.173 | 0.041 | 0.000 | 0.24 | 0.00 | 0.039 | ||||||||||
l-Lactic acid | 147 | 765 | –0.0097 | –0.0093 | 0.301 | 0.306 | 0.206 | 1.02 | 0.69 | 0.519 | ||||||||||
Succinic acid | 148 | 999 | –0.0241 | 0.0232 | 0.541 | 0.677 | 0.250 | 1.25 a | 0.46 b | 0.006 | ||||||||||
Pyruvic acid | 174 | 756 | –0.0010 | –0.0000 | 0.020 | 0.012 | 0.005 | 0.61 a | 0.27 b | <0.001 | ||||||||||
Glyceric acid | 189 | 1024 | 0.0025 | 0.0005 | 0.036 | 0.038 | 0.067 | 1.05 | 1.87 | 0.165 | ||||||||||
Pipecolic acid | 156 | 1051 | 0.0139 | 0.0068 | 0.144 | 0.171 | 0.275 | 1.18 | 1.90 | 0.263 | ||||||||||
Mucic acid | 333 | 1763 | 0.0003 | 0.0009 | 0.317 | 0.348 | 0.317 | 1.10 | 1.00 | 0.387 | ||||||||||
Citric acid | 347 | 1529 | –0.1140 | 0.0781 | 8.026 | 7.309 | 6.357 | 0.91 | 0.79 | 0.069 | ||||||||||
Isocitric acid | 273 | 1531 | 0.0023 | 0.0029 | 0.000 | 0.079 | 0.056 | Dark sp. | Dark sp. | 0.537 | ||||||||||
Citramalic acid | 147 | 1169 | 0.1520 | 0.1640 | 5.783 | 8.406 | 7.776 | 1.45 | 1.34 | 0.005 | ||||||||||
Maleic acid | 147 | 989 | –0.0018 | –0.0019 | 0.048 | 0.055 | 0.025 | 1.15 | 0.52 | 0.502 | ||||||||||
Fumaric acid | 245 | 1033 | –0.0020 | 0.0002 | 0.159 | 0.150 | 0.119 | 0.94 | 0.75 | 0.187 | ||||||||||
Glucaric acid | 333 | 1732 | –0.0017 | 0.0010 | 0.480 | 0.439 | 0.434 | 0.92 | 0.90 | 0.843 | ||||||||||
Gluconic acid | 333 | 1698 | –0.0079 | 0.0038 | 0.459 | 0.213 | 0.255 | 0.46 | 0.56 | 0.034 | ||||||||||
2-Hydroxycinnamic acid | 293 | 1397 | 0.0112 | –0.0026 | 0.668 | 0.884 | 0.777 | 1.32 | 1.16 | 0.298 | ||||||||||
4-Hydroxycinnamic acid | 293 | 1624 | –0.0011 | –0.0023 | 0.038 | 0.042 | 0.015 | 1.09 | 0.40 | 0.464 | ||||||||||
Citraconic acid | 147 | 1040 | –0.0000 | 0.0001 | 0.002 | 0.004 | 0.001 | 2.32 | 0.77 | 0.437 | ||||||||||
Glycolic acid | 147 | 775 | –0.0008 | 0.0002 | 0.023 | 0.012 | 0.008 | 0.53 | 0.35 | 0.021 | ||||||||||
Malonic acid | 147 | 890 | –0.0094 | 0.0005 | 0.736 | 0.732 | 0.671 | 1.00 | 0.91 | 0.926 | ||||||||||
d-Malic acid | 147 | 1187 | –0.3360 | 0.1160 | 5.901 | 5.240 | 1.798 | 0.89 a | 0.30 b | <0.001 | ||||||||||
Oxalic acid | 147 | 824 | –0.0130 | 0.0027 | 0.325 | 0.272 | 0.177 | 0.84 a | 0.54 b | 0.013 | ||||||||||
Polyols | ||||||||||||||||||||
Galactinol | 204 | 2708 | –0.0070 | –0.0037 | 0.092 | 0.031 | 0.010 | 0.33 | 0.11 | 0.101 | ||||||||||
Palatinitol | 361 | 2555 | 0.0011 | 0.0009 | 0.034 | 0.036 | 0.053 | 1.06 | 1.56 | 0.417 | ||||||||||
d-Mannitol | 319 | 1648 | 0.0471 | 0.1800 | 0.477 | 1.643 | 1.161 | 3.45 | 2.43 | 0.020 | ||||||||||
Pinitol | 260 | 1550 | –0.4390 | –0.0313 | 22.215 | 22.108 | 18.608 | 1.00 | 0.84 | 0.223 | ||||||||||
allo-Inositol | 318 | 1745 | –0.0008 | –0.0010 | 0.216 | 0.202 | 0.201 | 0.94 | 0.93 | 0.984 | ||||||||||
Phosphates | ||||||||||||||||||||
O-Phospho-l-threonine | 370 | 1534 | –0.0026 | –0.0008 | 0.037 | 0.000 | 0.000 | 0.00 | 0.00 | – | ||||||||||
Phosphoric acid | 299 | 967 | –0.0072 | 0.0056 | 3.422 | 2.796 | 3.168 | 0.82 | 0.93 | 0.245 | ||||||||||
Phosphoenolpyruvic acid | 369 | 1302 | 0.0014 | –0.0000 | 0.008 | 0.012 | 0.028 | 1.57 a | 3.68 b | 0.007 | ||||||||||
O-Phosphocolamine | 299 | 1485 | 0.0161 | –0.0036 | 0.010 | 0.089 | 0.203 | 8.79 a | 19.97 b | <0.001 | ||||||||||
Pyrophosphate | 451 | 1377 | –0.0013 | 0.0001 | 0.021 | 0.009 | 0.003 | 0.41 | 0.16 | <0.001 | ||||||||||
Adenosine-5-monophosphate | 315 | 2751 | –0.0188 | –0.0008 | 0.366 | 0.161 | 0.111 | 0.44 a | 0.30 b | <0.001 | ||||||||||
3-Phosphoglycerate | 357 | 1519 | 0.0015 | –0.0044 | 0.116 | 0.018 | 0.134 | 0.16 | 1.16 | 0.291 | ||||||||||
3-Phosphoglyceric acid | 387 | 1519 | 0.0028 | 0.0034 | 0.086 | 0.096 | 0.132 | 1.12 | 1.53 | 0.232 | ||||||||||
d-Glucose-6-phosphate | 387 | 2052 | –0.0221 | –0.0051 | 0.348 | 0.144 | 0.098 | 0.41 | 0.28 | 0.019 | ||||||||||
Uridine 5’-monophosphate | 315 | 2563 | –0.0029 | –0.0003 | 0.044 | 0.074 | 0.030 | 1.65 | 0.69 | 0.181 | ||||||||||
Glycerol 1-phosphate | 357 | 1471 | –0.0011 | –0.0011 | 0.155 | 0.151 | 0.126 | 0.97 | 0.82 | 0.556 | ||||||||||
Other metabolites | ||||||||||||||||||||
Putrescine | 174 | 1427 | –0.0014 | –0.0003 | 0.063 | 0.045 | 0.037 | 0.73 | 0.59 | 0.009 | ||||||||||
Spermidine | 174 | 1952 | –0.0010 | –0.0004 | 0.043 | 0.043 | 0.024 | 1.01 | 0.56 | 0.320 | ||||||||||
Uracil | 241 | 1021 | 0.0001 | –0.0004 | 0.021 | 0.009 | 0.015 | 0.45 | 0.73 | 0.619 | ||||||||||
Urea | 147 | 922 | 0.0009 | –0.0023 | 0.013 | 0.034 | 0.035 | 2.63 | 2.75 | 0.488 | ||||||||||
Adenine | 264 | 1555 | –0.0014 | 0.0010 | 0.019 | 0.025 | 0.013 | 1.32 | 0.65 | 0.440 | ||||||||||
Adenosine | 236 | 2342 | 0.0004 | 0.0066 | 0.028 | 0.111 | 0.053 | 3.96 a | 1.88 b | 0.007 | ||||||||||
Norvaline | 72 | 783 | 0.0004 | 0.0008 | 0.000 | 0.006 | 0.007 | Dark sp. | Dark sp. | 0.246 | ||||||||||
Loganin | 361 | 2539 | 0.0032 | 0.0032 | 0.254 | 0.290 | 0.287 | 1.14 | 1.13 | 0.557 | ||||||||||
Thymine | 255 | 1057 | 0.0003 | 0.0004 | 0.011 | 0.024 | 0.015 | 2.14 | 1.35 | 0.235 | ||||||||||
l-Norleucine | 158 | 960 | 0.0206 | 0.0045 | 0.014 | 0.163 | 0.226 | 11.39 | 15.79 | 0.040 | ||||||||||
Porphine | 285 | 1020 | 0.0013 | 0.0012 | 0.008 | 0.024 | 0.028 | 2.91 | 3.38 | 0.146 | ||||||||||
Trans-3-hydroxy-l-proline | 230 | 1221 | 0.0002 | –0.0001 | 0.000 | 0.001 | 0.002 | Dark sp. | Dark sp. | 0.186 | ||||||||||
Methyl-β-d-galactopyranoside | 204 | 1699 | –0.0177 | –0.0082 | 0.482 | 0.258 | 0.229 | 0.54 | 0.48 | <0.001 | ||||||||||
N-methyl-d-glutamic acid | 260 | 1382 | –0.0086 | –0.0006 | 0.110 | 0.197 | 0.033 | 1.78 a | 0.30 b | 0.009 | ||||||||||
1-Methyl-nicotinamide | 179 | 1155 | –0.0009 | –0.0004 | 0.015 | 0.005 | 0.002 | 0.31 | 0.11 | 0.011 |
For each identified metabolite, GC-MS characteristics (fragment mass, RI, retention index), loadings in principal components of Fig. 3, average response (resulting from five biological replications), and response ratios of each dark period to the normal photoperiod are presented.
Response ratios representing significant differences (P < 0.05) are indicated in bold, while significant differences between the relative responses measured after 24h and 72h darkness are indicated by different letters. The P-value represents the level of significance between the relative responses of each compound (ANOVA), when at least two average responses are >0.