Table 1.
Efficiency and nucleotide sequences of primers used in qPCR
| Transcript* | Calculated primer efficiency | Forward Primer Sequence | Reverse Primer Sequence | Fragment size (bp) |
|---|---|---|---|---|
| β-actin | 1.91 | 5'-CCAGGCTGTTCAGTCTCTGTAT-3' | 5'-CGCTCGGTAAGGATCTTCATCA-3' | 180 |
| ADS | 1.88 | 5'-GGGAGATCAGTTTCTCATCTATGAA-3' | 5'-CTTTTAGTAGTTGCCGCACTTCTT-3' | 95 |
| ALDH1 | 1.82 | 5'-CATCGGAGTAGTTGGTCACAT-3' | 5'-GTTTCTGACCCAAATCCAGGTTGA-3' | 120 |
| CPR | 1.90 | 5'-GCTCGGAACAGCCATCTTATTCTT-3' | 5'-GAAGCCTTCTGAGTCATCTTGTGT-3' | 174 |
| CPS | 1.85 | 5'-CAACGATGTAGAAGGCTTGCTTGA-3' | 5'-GTAGATAGTGTTGGGTTGGTGTGA-3' | 150 |
| CYP71AV1 | 1.83 | 5'-CGAGACTTTAACTGGTGAGATTGT-3' | 5'-CGAAGCGACTGAAATGACTTTACT-3' | 144 |
| DBR2 | 1.91 | 5'-GCGGTGGTTACACTAGAGAACTT-3' | 5'-ATAATCAAAACTAGAGGAGTGACCC-3' | 228 |
| DBR2/OPR3 | 1.84 | 5'-ATCATCAACAAGCAAGCCCATTTCAAA-3' | 5'-CGATAGTCTTCAACCACCTCTAGTA-3' | 125 |
| DXR | 1.92 | 5'-GGTGATGAAGGTGTTGTTGAGGTT-3' | 5'-AGGGACCGCCAGCAATTAAGGT-3' | 160 |
| DXS | 1.92 | 5'-GTGCTTCCAGACCGTTACATTGA-3' | 5'-AGCCTCTCGTGTTTGCCCAAGGT-3' | 120 |
| ECS | 1.86 | 5'-GCAACAAGCCTACGAATCACTCAA-3' | 5'-CGTGAAAAATTAAGGACCCTCATAG-3' | 126 |
| FDS1/FDS2 | 1.93 | 5'-ATCTGCCCTTGGTTGGTGTATTGA-3' | 5'-GTTGCCCTCTGCGTGTATGAGA-3' | 92 |
| GAS | 1.83 | 5'-CTCGTTACTCCTTGGCAAGAATCAT-3' | 5'-GCTCCATAGCACTAATATCCCACTT-3' | 147 |
| HDR | 1.86 | 5'-TCAGGAGCGACAAGATGCTATGTA-3' | 5'-AGTGTGAGGTGTTGCTTGAGTTGA-3' | 95 |
| HMGR | 1.83 | 5'-GGGCGTGGAAAATCTGTTGTGTTC-3' | 5'-GAACCAGCAATAGCAGAACCAGTAA-3' | 136 |
| PAL | 1.87 | 5'-ATCGGGAAACTCATGTTCGCTCAA-3' | 5'-AACTTGGGTTACGTCCACCAGAAA-3' | 97 |
| RED1 | 1.92 | 5'-TGTCAACTGTGTCCATCCAGGTT-3' | 5'-ACCATCATCGGGCAACAAAGCAA-3' | 118 |
| SQS | 1.82 | 5'-GACCAGTTCCACCATGTTTCTACT-3' | 5'-GCTTTGACAACCCTATTCCAACAAG-3' | 190 |
The mean efficiency of the amplicons was calculated by the program Linreg v. 12.1 based on the log linear phase of the amplification curve. The transcripts in bold were used as reference genes in the calculation of expression ratios.
* ADS, amorpha-4,11-diene synthase; ALDH1, aldehyde dehydrogenase 1; CPR, cytochrome P450 reductase; CPS, β-caryophyllene synthase; CYP71AV1, amorphadiene-12-hydroxylase; DBR2, artemisinic aldehyde Δ11(13) reductase; DXR, 1-deoxy-D-xylulose 5-phosphate reductase; DXS, 1-deoxy-D-xylulose 5-phosphate synthase; ECS, epi-cedrol synthase; FDS, farnesyl diphosphate synthase; GAS, germacrene A synthase; HDR, hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase; HMGR, 3-hydroxy-3-methyl-glutaryl-CoA reductase; OPR3, 12-oxophytodienoate reductase; PAL, phenylalanine ammonia lyase; RED1, dihydroartemisinic aldehyde reductase; SQS, squalene synthase.