Table 1. Metabolic Profiles of the gbp1 Mutant.
| Biochemical Family | Metabolite | Fruit Developmental Stage | Analytical Method | MSI Levela | NMR or MS Signalsb | |||
|---|---|---|---|---|---|---|---|---|
| 15 DPA | 20 DPA | 25 DPA | Red Ripe | |||||
| Amino Acids | Alanine | — | — | WT* | — | NMR | MSI1 | B1.4762; B1.4907 |
| Asparagine | — | — | WT** | — | NMR | MSI1 | B2.7978; B2.8135; B2.8461; B2.8525; B2.9398 | |
| Phenylalanine | — | — | WT** | — | NMR | MSI1 | B7.3342; B7.3395 | |
| MS(pos) | MSI1 | X41 | ||||||
| Valine | — | WT** | WT** | - | NMR | MSI1 | B0.9991; B1.0115; B1.0503 | |
| Glutamine | — | WT** | WT** | WT* | NMR | MSI1 | B2.4369;B2.4451;B2.4515;B2.4569;B2.4623;B2.4689;B2.4778 | |
| Aspartic acid | — | WT* | WT* | gbp1* | NMR | MSI1 | B2.6126;B2.6484;B2.6642;B2.7853;B2.7931;B2.8214;B2.8274 | |
| Isoleucine | — | WT* | WT* | gbp1* | NMR | MSI1 | B1.0175;B1.0304 | |
| Glutamic acid | — | gbp1* | gbp1* | - | NMR | MSI1 | B2.0372;B2.0520 | |
| GABA | — | gbp1* | gbp1** | gbp1** | NMR | MSI1 | B1.8691;B1.8879;B1.9021;B1.9168;B1.9311;B2.2887;B2.3051;B2.3202;B3.0264 | |
| Tyrosine | — | — | gbp1** | gbp1** | NMR | MSI1 | B6.8424;B6.8598;B7.1731;B7.1915 | |
| Organic Acids | Fumaric acid | — | — | gbp1** | gbp1** | NMR | MSI1 | B6.5291 |
| Lactic acid | — | — | — | gbp1** | NMR | MSI3 | B1.3608 | |
| Isobutyric acid | — | — | — | gbp1* | NMR | MSI3 | B1.0840; B1.0964 | |
| Pantothenic acid hexose | — | — | — | gbp1* | MS(neg) | MSI3 | X852 | |
| Alcohols | Ethanol | — | — | — | gbp1** | NMR | MSI1 | B1.1753 |
| Sugars | Glc | — | WT* | WT* | WT* | NMR | MSI1 | B3.1862; B3.2052; B3.6791; B3.7049; B3.7252; B3.7354; B3.8619;B4.5769; B4.5985; B5.1805; B5.1996 |
| Fru | — | — | gbp1* | WT* | NMR | MSI1 | B3.7692; B3.9612 | |
| Gal | — | — | gbp1* | WT* | NMR | MSI1 | B4.5262; B5.2358 | |
| Suc | — | — | — | gbp1** | NMR | MSI1 | B4.1662; B4.1847; B5.3988; B5.4131 | |
| Cyclitols and derivatives | Inositol | — | — | gbp1* | — | NMR | MSI1 | B3.2454; B3.2621 |
| Alkaloids | Trigonelline | — | — | — | gbp1* | NMR | MSI1 | B4.4496; B8.8670; B9.1435 |
| Glycoalkaloids | Deglucosylated lycoperoside F/deglucosylated esculeoside A | — | — | — | gbp1** | MS(pos) | MSI3 | X1667 |
| Lycoperoside C | — | — | gbp1** | MS(pos) | MSI3 | X1661 | ||
| Lycoperoside F-G/esculeoside A isomer 2 | — | — | — | gbp1* | MS(pos) | MSI3 | X1692 | |
| Alpha-Tomatine | — | — | — | gbp1* | MS(pos) | MSI3 | X1638 | |
| Dehydrotomatine | — | — | — | gbp1** | MS(pos) | MSI3 | X1634 | |
| Hydroxycinnamic acid derivatives | Chlorogenic acid (3CQA) | — | gbp1** | gbp1** | gbp1** | NMR | MSI1 | B1.9856;B2.0063;B4.1933;B4.1990;B4.2059;B4.2125;B6.3635;B6.3963;B6.8889;B7.0665;B7.0737;B7.084;B7.0912;B7.1545;B7.1620;B7.6106 |
| — | — | gbp1** | — | MS(pos) | MSI2 | X492 | ||
| — | — | — | gbp1** | MS(neg) | MSI2 | X661 | ||
| Cryptochlorogenic acid (4CQA) | — | — | — | gbp1* | MS(neg) | MSI2 | X663 | |
| Dicaffeoylquinic acid isomer 2 | — | — | — | gbp1** | MS(pos) | MSI3 | X982 | |
| MS(neg) | MSI3 | X1893 | ||||||
| Dicaffeoylquinic acid isomer 1 | — | — | — | gbp1** | MS(neg) | MSI3 | X1895 | |
| Tricaffeoylquinic acid | — | — | — | gbp1** | MS(neg) | MSI3 | X2798 | |
| — | — | — | MS(pos) | MSI3 | X1397 | |||
| Feruloylputrescine isomer 2 | — | — | — | gbp1** | MS(pos) | MSI3 | X214 | |
| Feruloylquinic acid | — | — | — | gbp1** | MS(neg) | MSI3 | X771 | |
| gbp1* | MS(pos) | MSI3 | X534 | |||||
| Feruloyltyramine | — | — | — | gbp1** | MS(neg) | MSI3 | X424 | |
| gbp1** | MS(pos) | MSI2 | X367 | |||||
| Coumaroylquinic acid isomer 1 | — | — | — | gbp1** | MS(neg) | MSI3 | X549 | |
| gbp1** | MS(pos) | MSI3 | X443 | |||||
| Coumaroylquinic acid isomer 2 | — | — | — | gbp1** | MS(neg) | MSI3 | X548 | |
| Flavonoids | Rutin | — | WT* | WT** | gbp1* | NMR | MSI3 | B1.3288;B1.3416,B6.5544;B6.5599;B6.9886;B7.0071;B7.6804;B7.6869 |
| Xanthosine | — | — | — | gbp1** | MS(neg) | MSI2 | X291 | |
| Inosine | — | — | — | gbp1** | MS(neg) | MSI3 | X225 | |
| Uridine monophosphate | — | — | — | WT* | MS(neg) | MSI2 | X472 | |
Annotated metabolites with significant differences (Wilcoxon test, FDR-corrected P-value < 0.05) in levels between the mutant and the wild type (WT) at one or several stages of fruit development are shown. The genotype (wild type or gbp1-c) with higher content in the pericarp is indicated (FC > 1.2 and > 2 are indicated by * and **, respectively). Only metabolites that are consistent between the two CRISPR lines are shown. Corresponding analytical methods and signals of detection are shown. Dashes indicate no significant difference.
Groups of the Metabolomics Standards Initiative (MSI) level according to Sumner et al. (2007). For NMR MSI1, 1D and 2D 1H and/or 13C confirmation + internal NMR database of reference compounds acquired in the same conditions of solvent and pH or spiking have been performed; for NMR MSI3, as only 1D 1H detection + external NMR database of reference compounds or literature data have been performed, annotations are putative.
NMR signals correspond to integrated spectral region or bucket centered at a chemical shift expressed in ppm (B1.4762, where “B” indicates Bucket and 1.4762 is the center of the bucket in ppm); MS signals correspond to software MS-DIAL variable IDs.