Table 6.
Detailed information of primers for 17 phenolic compound biosynthesis pathway genes for real-time quantitative gene expression analysis (qRT-PCR).
| Gene | Probe set IDa | Chrb | Primer* | Sequence (5′-3′) | Tm (°C) | Amplicon size (bp) |
|---|---|---|---|---|---|---|
| Phenylalanine ammonia lyase (PAL 5) | Ta.28046.1.A1_at | 2BL | F | AGGAGCGCCGTGGAGAATGGCACT | 625 | 141 |
| R | CTCTTCGCCAGGAGACCGCGTCTT | 625 | ||||
| Cinnamate 4-hydroxylase (C4H1) | Ta.9643.2.S1_x_at | 3B | F | CATCCCGCTGCTGGTGCCCCA | 625 | 123 |
| R | TCCACTGCTCCGGGTTGTTGGCGA | 625 | ||||
| 4-Coumarate coenzyme A ligase (4CL) | Ta.5623.1.S1_x_at | 6AS | F | CTACAAGAGGATCCACAAGGT | 524 | 159 |
| R | TGCTGGCTGGCTGAGTGCCTG | 602 | ||||
| p-coumaroyl quinate/shikimate-3-hydroxylase (C3′H) | Ta.20769.1.S1_x_at | 1AL | F | TCGCTGCTCCCGTTCGCCATC | 602 | 134 |
| R | GGCTTGATCTGCCGCAGGTTG | 583 | ||||
| Caffeoyl-CoA O-methyltransferase 1 (CCoMT1) | Ta.18653.1.S1_at | 4AL | F | TGCTGATCAAGATGGCGGGCG | 583 | 129 |
| R | ACTCGCGGTCGGTGTCGATGG | 602 | ||||
| Cinnamoyl-CoA reductase (CCR) | Ta.13990.1.S1_at | 5BL | F | TATACGAGACGGTGAAGAGCCTCC | 591 | 180 |
| R | GTCCCATAGAATTAACTAGCCGACAGC | 597 | ||||
| Cinnamyl alcohol dehydrogenase (CAD) | Ta.13798.1.S1_at | 6AS | F | GGTGCTCCAGTTCTGCGTCGACAA | 608 | 157 |
| R | TCAGGCGGCGTCCTCGATGTT | 583 | ||||
| Ferulate 5-hydroxylase 1 (F5H1) | Ta.Affx108591.1.S1_x_at | 1BL | F | ACACCGTCGAATCGAACACCCAAG | 591 | 147 |
| R | CGAAGACCACCGAGGCGACGAACA | 625 | ||||
| Caffeic acid -O-methyl transferase (COMT) | Ta.336.2.S1_at | 7DL | F | AAGAACGCCATCGAGCTGGGTCTC | 608 | 180 |
| R | CTCCTCCATCGTGCACGACACCAC | 625 | ||||
| Chalcone synthase (CHS) | Ta.9172.1.S1_at | 2AL | F | CTTCCACCTGCTCAAGGACGTGCC | 625 | 267 |
| R | GAGCGCTTGCGCATCTCATCCATG | 608 | ||||
| Flavonol synthase1 (FLS1) | Ta.25327.1.A1_at | 6AL | F | GGTGTTCAAGGACGGATGCTGGTA | 591 | 157 |
| R | CGGCCACGACATCCGCGTCTT | 602 | ||||
| Flavone synthase (FNS) | Ta.9332.1.S1_x_at | 5DL | F | TGATGGGCAAGAAGTACCTGGAGA | 574 | 166 |
| R | TACCCCTGCAGGTCCATCCAG | 583 | ||||
| 1,2 Rhamnosyl transferase (1,2 RhaT) | Ta.30441.1.A1_at | 2BS | F | CGGCCGTTCTTCGTCGTGCTCAA | 606 | 134 |
| R | GCTGCTGCACCCACCCCGTGT | 622 | ||||
| Benzoate 4-monoxygenase (B4MO) | Ta.254.1.S1_s_at | 6DL | F | CGCCGAGGACATGCACGAGTG | 602 | 199 |
| R | GAAGACGCCGTGCCAGAAGGG | 602 | ||||
| Quercetin-3-O-glucoside L–Rhamnosyl transferase (Q3-GRhaT) | Ta.Affx139860.1.A1_at | 3B | F | GGCACGCGGGATGGTGGTTTC | 602 | 142 |
| R | GGTCCAGCTCAGCATCGTCAC | 583 | ||||
| Vanillin dehydrogenase (VDH) | Ta.2955.2.A1_at | 7DL | F | ACTACATCGAGCCCACGGTTTTCA | 574 | 140 |
| R | GTACCTCGTGTCGTTGGCCCT | 642 | ||||
| Vanillin synthase (VS) | Ta.Affx107168.1.S1_x_at | 5BL | F | CTTCCATCCCAGGCATTTGAGTAC | 574 | 216 |
| R | GATCACCTCAAAGGCAACACTCAC | 574 |
a
The primer pairs were designed using the probe sets of GeneChip®; Wheat Genome Array (Affymetrix, Santa Clara, USA) corresponding to eighteen phenylpropanoid pathway genes.
b
Chromosome (Chr) mapping was done using IWGSC's wheat genome sequences;
*
F, forward and R, reverse primer.