TABLE 1.
Primers used in this studya
| Name | Sequence (5′–3′) | Purpose |
|---|---|---|
| lcc9-antisense1-fwd | CTCCCATCTACACACAACAAGCTTATCGCCCGGGATTCTCATAGTTGTAAGTGCTGC | Cloning of lcc9 antisense fragment 1 |
| lcc9- antisense 1-rev | CACTGGCCCTCTGGTCAACTATAATATTATAGATGGGCCTTGGACCTGCCG | Cloning of lcc9 antisense fragment 1 |
| lcc9- antisense 2-fwd | CTCCCATCTACACACAACAAGCTTATCGCCCGGACCACTTCCTCCTGGGGCA | Cloning of lcc9 antisense fragment 2 |
| lcc9- antisense 2-rev | CACTGGCCCTCTGGTCAACTATAATATTATCTCTCATGGTCGACGAAATCCAGATC | Cloning of lcc9 antisense fragment 2 |
| PF | ACATCCACCATCTCCGTTTTCTCCCAT | PCR of co-transformants |
| PR | TGACTATAGCAGCCTCCTACCACTG | PCR of co-transformants |
| qRT-13048-F | CTCTGGAGTTATGGTAGGAATGGGC | qRT-PCR of β-actin |
| qRT-13048-R | GATGCCATGTTCGATGGGGTACTTG | qRT-PCR of β-actin |
| qRT-00146-F | CCGCCAACAACTATGCCCGTGGTCA | qRT-PCR of α-tubulin |
| qRT-00146-R | TCCTCCACCGAAGGAGTGGAAGACG | qRT-PCR of α-tubulin |
| qRT-10735-F | TTCGCTCCTGGTGACGCTTCTA | qRT-PCR of cyt-c |
| qRT-10735-R | GCCGAAGATGCCGAAGAGGTTG | qRT-PCR of cyt-c |
| qRT-11833-F | AAGGCGAAGATTCAGGACAAGGAAGGTA | qRT-PCR of ubc |
| qRT-11833-R | CGAAGGCGGAGAACCAAGTGAAGAGTA | qRT-PCR of ubc |
| qRT-07869-F | GAAGAAGACTCTCACTCCGTCAACG | qRT-PCR of capzb |
| qRT-07869-R | CTGTGTGGTGAGCTGGAGCATGA | qRT-PCR of capzb |
| qRT-lcc9-F | ATGTCCAGGAAACTTTTCTCTCTCG | qRT-PCR of lcc9 |
| qRT-lcc9-R | ATGTTCGAGACCGTCATGGTACT | qRT-PCR of lcc9 |
| qRT-07201-F | AGCGCAAGCTGACCCCTTGGT | qRT-PCR of sga1 |
| qRT-07201-R | ACAAGTCCAGCCGCTGCCCC | qRT-PCR of sga1 |
| qRT-05652-F | CAAGGATCGACGGAGGGCGCA | qRT-PCR of tkl |
| qRT-05652-R | TCCACTTCCGAGCCGCCACC | qRT-PCR of tkl |
| qRT-02494-F | AGCAAGGTGGATGTCCACGAGTCC | qRT-PCR of vwfa |
| qRT-02494-R | GGCCACTGCCAGCGACTTGGT | qRT-PCR of vwfa |
| qRT-06452-F | TACGCTGGAATGGACGCCGGA | qRT-PCR of exg |
| qRT-06452-R | TGCCGCGGGCCCAAAAGTTCTTC | qRT-PCR of exg |
| qRT-03564-F | CACCTTCGCTTGTTCTACGGATCCA | qRT-PCR of cyp98A3 |
| qRT-03564-R | TCAACCCAGAAGCGTCCGGGAATAC | qRT-PCR of cyp98A3 |
| qRT-05291-F | CTAAGCAGCATAATGTGAAAGCCCT | qRT-PCR of Chi |
| qRT-05291-R | GGAAATTCCCAGTCAAAGTCGATACCA | qRT-PCR of Chi |
| qRT-11387-F | ATCGAGATGATCGAGCATGTCGGC | qRT-PCR of cfa |
| qRT-11387-R | TGATGACAGCGGCGGCCTTTTG | qRT-PCR of cfa |
| qRT-08324-F | TGCAGAGGAAGAGCGACCTCGGA | qRT-PCR of samB |
| qRT-08324-R | CCAGATGCTGAGAGGGAAACGGAGG | qRT-PCR of samB |
| qRT-09241-F | CCGGATTTGAAGACGATCAAGAGCTT | qRT-PCR of psd |
| qRT-09241-R | GAGAAGGTACATGGGAACGCCGATAG | qRT-PCR of psd |
| qRT-09422-F | TTATCGCATACTCGCCCCTCGGA | qRT-PCR of akr |
| qRT-09422-R | AAGGCATCGACGATTGCCATGTTG | qRT-PCR of akr |
| qRT-14829-F | TGCTGGAGAGCAAGGTTGTGTCTCA | qRT-PCR of gst |
| qRT-14829-R | AAGGGAATGGTGTAGTGGGGCGAGC | qRT-PCR of gst |
| qRT-05005-F | GCCATCCGCTCCAGTGCCTTCAAT | qRT-PCR of cgl-II |
| qRT-05005-R | GCGGATGCTGATGTGGAGGAGGATG | qRT-PCR of cgl-II |
| qRT-02499-F | CAGTCAGAGAGTTGCCTATACTCTCA | qRT-PCR of tdo |
| qRT-02499-R | GGATGATGGTGGCAGAGAGTGAATG | qRT-PCR of tdo |
| qRT-07229-F | CATCTTAGCTGGCTCGGCTTCTTC | qRT-PCR of nrt |
| qRT-07229-R | ACGATGTTGGAGTTGGCGACTTCG | qRT-PCR of nrt |
| qRT-lcc1-F | ATGTTCAAGAACCTCCTCTCGT | qRT-PCR of lcc1 |
| qRT-lcc1-R | ACGTTCGCGTTGGTGAGGGTCAT | qRT-PCR of lcc1 |
| qRT-lcc5-F | ATGTCGTTTGCTTGGAAAGCAT | qRT-PCR of lcc5 |
| qRT-lcc5-R | TGGTCATAGTATCTTGGTTGCCAAT | qRT-PCR of lcc5 |
| qRT-08281-F | TGTACAAGATGCTCGAAGACCCA | qRT-PCR of skn7 |
| qRT-08281-R | TTCATGTCCTTTACGACGAAACAGTC | qRT-PCR of skn7 |
a
Five candidate reference genes chosen were β-actin, capzb (F-actin capping protein beta subunit), α-tub (tubulin), cyt-c (cytochrome c), and ubc (ubiquitin). Fifteen randomly chosen genes for qRT-PCR to validate gene expression data in the RNA-seq experiment were lcc9 (laccase 9), sga1 (glucoamylase), tkl (TKL/TKL-ccin protein kinase), vwfa (VWFA domain protein), exg (glucan 1, 3-beta-glucosidase), cyp98A3 (cytochrome P450 98A3), Chi (endochitinase), cfa (cyclopropane-fatty-acyl-phospholipid synthase), samB (MYND-type Zinc finger protein), psd (phosphatidylserine decarboxylase), akr (aldo/keto reductase), gst (glutathione S-transferase), cgl-II (galectin-2), tdo (tryptophan 2,3-dioxygenase), and nrt (nitrate transporter).