Table 4 Genes of interest and their encoding enzymes selected for gene expression analysis.
Metabolites | Enzyme | Abbreviation | EC number | Gene NCBI accession number | Primer sequence | Amplicon length (bp) | Ta (°C) | Tm (°C) | Amplification efficiency |
|---|---|---|---|---|---|---|---|---|---|
Caffeic acid | Caffeic acid 3-O-methyltransferase | COMT | EC 2.1.1.68 | XM_003634113.2 | Fw: GTATGACCCCAACAACTATC | 88 | 60 | 78.4 | 1.88 ± 0.02 |
Rev: GACCATGGGGAGAACTGA | |||||||||
Catechin | Leucoanthocyanidin reductase 2 | LAR2 | EC 1.17.1.3 | NM_001281160.1 | Fw: TGTAACCGTGGAAGAAGATGA | 92 | 60 | 80.6 | 1.88 ± 0.02 |
Rev: ATGAAGATGTCGTGAGTGAAG | |||||||||
Epicatechin | Anthocyanidin reductase | ANR | EC 1.3.1.77 | NM_001280956.1 | Fw: ATCAAGCCAGCAATTCAAGGA | 93 | 60 | 76.2 | 1.88 ± 0.006 |
Rev: CAGCTGCAGAGGATGTCAAA | |||||||||
Dodecanoic acid/hexadecanoic acid | Palmitoyl-acyl carrier protein thioesterase B | FatB | EC 3.1.2.21/3.1.2.14 | XM_019223124.1 | Fw: TCGCAAACCCTAGAAACCAAT | 112 | 60 | 77.5 | 1.93 ± 0.05 |
Rev: AATGAGGGAAGGAGGAAAATG | |||||||||
Myo inositol | Myo-inositol monophosphatasea | IMPL1 | EC 3.1.3.25 | XM_002276661.3 | Fw: ATCCCAAACGCTACCCAAAAA | 119 | 60 | 80.9 | 1.96 ± 0.02 |
Rev: TAACAGCTTCCATCACAACCT | |||||||||
Quercetin/dihydroquercetin | Flavonoid 3′,5′-hydroxylaseb | F3′5′H | EC 1.14.14.81 | XM_003632164.3 | Fw: GTGGTGCCGGAGATGTTA | 173 | 56 | 80.1 | 1.83 ± 0.05 |
Rev: TGCGATGGACGGAATAAAAT | |||||||||
Quercetin-3-O-glucoside (Isoquercitrin) | UDP-glucose:flavonoid 3-O-glucosyltransferase | UFGT | EC 2.4.1.91 | AF000372.1 | Fw: AGGGGATGGTAATGGCTGT | 151 | 60 | 84.7 | 1.97 ± 0.01 |
Rev: ATGGGTGGAGAGTGAGTTAG |
