Genetic Resources and Crop Evolution

TaqSH1-D, wheat ortholog of rice seed shattering gene qSH1, maps to the interval of a rachis fragility QTL on chromosome 3DL

Mazen Katkout1, Shun Sakuma1, Kanako Kawaura1, Yasunari Ogihara1,*

1 Kihara Institute for Biological Research and Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, Maioka-cho 641-12, Totsuka-ku, Yokohama 244-0813, Japan

* Corresponding author. E-mail:

Table S1 Primers used for gene isolation and sequencing from both parental lines, Chinese Spring (CS) and the synthetic wheat line S-6214
Primer name / 5′-Sequence-3′ / Amplicon size
For PCR
WBLH4-1/F / ACAGCAGCGGAGACAGAG / 2536 bp
TaqSH1-D-1669/R / ACGGCCGATCTTATTAGCTCC
WBLH4b/F / CAACACCAGCAAGATGCCG / 1924 bp
WBLH4b/R / CATTGGCTTCCAGAGCCG
Cg352b/F / TGGTGCTAGACTGATAAAAGC / 2176 bp in CS
Cg3615/R / TTTGGGGCACTGTGGCATTA / 1987 bp in S-6214
Cg3615/F / TCCTTGTTTATGCCCAAAAGTTT / 1148 bp
Cg2717/R / TATGGCCATGAATAAACAGC
For sequencing
WBLH4-317/F / GCTTCATAACCATGCACACCAC
WBLH4-310/R / GCATGGTTATGAAGCAGCATG
WBLH4-811/F / AGGAGATCTGCGACGTGG
WBLH4-811/R / CCACGTCGCAGATCTCCT
Cg6717/F / CATCCACATGAAATCCAAAGCTC
Cg6717/R / GAGCTTTGGATTTCATGTGGATG
Cscg1/R / AAGTGCTCGAAGAGCCAAGC
wblh4b-w/F / GGCCTGGACCTCTAGTTGTT
wblh4b-w/R / GGATGTAATAGGCAAAACAACCA
cg352b/R / AGGATACCTGCAAAGTTTCAA
Syncg1/F / GGCTCACCAGCACTCGTC
Syncg2/R / CGAACGAGCCCATCACGT
Cg2717/F / CGAGCCTGTCTGAGCTACC

1 ACAGCAGCGGAGACAGAGGGAGATACCACCAGCACCAGGCCAGCCAGCCAGGCCCGGCGC 60

61 TCGGAAAGTGCGGAGCCGTTTCCCCTCCACCTCCACTCTCTCCAGGGCCACCCACCCAGC 120

121 CCCAGCCCCAGCCCAGCCAGGTGCTCCTGCTCCTCCTGCTCTACCTCGCTGCCCGCCACC 180

181 CACCGCGTACGTACGTGCGGTGCGGGCGCTCGCCATGTCGTCTCCCGCCGGCGGGTACGG 240

1 M S S P A G G Y G 9

241 CGGCGCCGAGGCCCACCACCACCAGCAGCACCACGGCCACATGCTGCTTCATAACCATGC 300

10 G A E A H H H Q Q H H G H M L L H N H A 29

301 ACACCACATGGCGACCGCGGCCGCCGCGGCGTCGGGCGGGCAGCTCTACCACGTGCCGCA 360

30 H H M A T A A A A A S G G Q L Y H V P Q 49

361 GCACAGCCGGCGCGAGAAGCTCCGGTTCCCGCCGGACGCCGCCGCGGAGGACTCGCCACC 420

50 H S R R E K L R F P P D A A A E D S P P 69

421 GACCCCTCTCGCCCACCACCACCAGCAGCAGCACCAGCAGCACCAGGCCGGGGCGTGGCC 480

70 T P L A H H H Q Q Q H Q Q H Q A G A W P 89

481 GCCCCCGGCCTTCTACTCCTACGCGTCCTCCTCCTCCTCCTACTCGCCGCACAGCCCGAC 540

90 P P A F Y S Y A S S S S S Y S P H S P T 109

541 GGTGCCGCAGGGCCAGCAGCTGGTGCTCAACGGGCTCACCGCCCAGCAGGTCACCGCGCA 600

110 V P Q G Q Q L V L N G L T A Q Q V T A Q 129

601 GCAGTTCCCGCAGATCCCCACGCAGAACTTCTCGCTCTCCCTCTCCTCCGCCTCGTCCAA 660

130 Q F P Q I P T Q N F S L S L S S A S S N 149

661 CCCCGCCACCACGCCCCCGGCGCCCAGGAAGCAGGAGCCAGGAGCCGCCGCGCCGGCCGC 720

150 P A T T P P A P R K Q E P G A A A P A A 169

721 GTGGCCGTACGGGCCCTTCACCGGCTACGCCACGGTGCTCGGGCGATCCAGGTTCCTCGG 780

170 W P Y G P F T G Y A T V L G R S R F L G 189

SKY domain

781 CCCGGCGCAGAAGCTGCTCGAGGAGATCTGCGACGTGGGGGGCGCGGCCGCGCACGCCGA 840

190 P A Q K L L E E I C D V G G A A A H A D 209

841 CACCAGCGTCCCGGACGAGGGCCCGCTCGACGCGGACGCCATGGACGGCGCCGACGACGC 900

210 T S V P D E G P L D A D A M D G A D D A 229

901 GGCCGGCCACGAGCTGGACACCTCCGGCCCCATGTCCGGCGCCGAGCAGCAGTGGAAGAA 960

230 A G H E L D T S G P M S G A E Q Q W K K 249

BEL domain

961 GACGAGGCTCATCTCCATGATGGAAGAGGTGAGCTAGCCACGACGCCGCTCGCTCCAAAT 1020

250 T R L I S M M E E V 259

1021 TTCCGACTTTTTCCGCCTCGTTCGCTTCGACGCCGCCACTGCAATTCTCCGGCTGGCTCC 1080

1081 TCGGCCGGGAGCGCGAAGAATCTTGCACATAGCGGTCACATCGCCAGCGACGCGGCCGTG 1140

1141 GCTTTGCCTGCTTTTATTCACTGTGCTTCCGCGGTTGCATGTGCTTTTGATTTGATGATC 1200

1201 ATTTCCATTGCTGCTGTTACATTTGCTCGGATTTTCCTTTTCTCCTTTCTCCACAGTGCC 1260

1261 GCACTGTTGGCGCCCAACAATTTGCTGCTCGCCTTTTTTTCTCATCAGAGGATCAAAGAT 1320

1321 TGGATGTTGATTAGTTTCACCGCACTAAGCACACCTTTTGCAAATATCATCTGCTACATG 1380

1381 AAACCACAACTGATTAGCAGCGTCCTCGAACGCTTTGGGAGGGTAGGTGATGCTGGAAAC 1440

1441 TAGTAGCAAAACTTTGGCCTGTCATGTCAAGAAAACCGTGTTGTTTCATCTCTCACATGG 1500

1501 TTAGGATAGGAGTACTTGACCCTCCCGCAGCGTCCTCTTCCAGCCTCTGGGGCAATACTG 1560

1561 TACATAACATGGGAGTCTACGTACGATGTAGAAAGCTTCCATTTATGGAGCTCTCAGTCT 1620

1621 GAGTCTCAGCCTCAGTAGGCGAAGTTGCTTGTCTGACTTCTTTATTAATGGCGTCTTACT 1680

1681 TCTATAGTTGGGAAGTAGGCTTTGCTTTGTTCATGTCACATCCACATGAAATCCAAAGCT 1740

1741 CCCAGCAAACAGCAAAGAAAATCTTTCCCATCTTCATGAGTAGTAGCTGCTTATCTCCTC 1800

1801 TAATCAAGTACCGTACTCACTTACCAATCCAGCTGCAATTAACCTCTCATCTCATTCTAC 1860

1861 TTCTAGGTGTGCAAGAGGTACCGGCAGTACTACCAGCAGGTCCAGGCCGTGATCGCGTCG 1920

260 C K R Y R Q Y Y Q Q V Q A V I A S 276

BEL domain

1921 TTCGAGAGCGTGGCTGGGTTCAGCAACGCCGCCCCGTTCACGGCGCTGGCGCTGAGGGTG 1980

277 F E S V A G F S N A A P F T A L A L R V 296

1981 ATGGCCAGGCACTTCAGGTGCATCAAGGGCATGATACTCAGCCAGCTGCGCAACACCAGC 2040

297 M A R H F R C I K G M I L S Q L R N T S 316

2041 AAGATGCCGGTCAAGGAAGGCATGAGCAAGGACATCACCATCTTCGGCCTCGGCGGCGGC 2100

317 K M P V K E G M S K D I T I F G L G G G 336

2101 GGCGGCGCCCCCGTCGGCGGCTTCCAGAGGGGTGGCAGCGTGAACGGCTTCGGCCAGCCG 2160

337 G G A P V G G F Q R G G S V N G F G Q P 356

2161 CACAACATCTGGCGCCCCCAGAGGGGCCTCCCCGAGCGCTCCGTCACGGTCCTCCGGGCT 2220

357 H N I W R P Q R G L P E R S V T V L R A 376

Homeodomain

2221 TGGCTCTTCGAGCACTTCCTGCACCCGTAAGTTGTTTTACTACCATCTGCTTATGGTAAT 2280

377 W L F E H F L H P Y 386

2281 TAGCATTAGCACTAGACGACGTACGTACTCTGCGCCTTTCCAAGATAATAATGTGTCTTT 2340

2341 GCGGTGCTACATCAACATGATTATGCTGGGCCTGGACCTCTAGTTGTTTTAGCTGATGGC 2400

2401 AAGCAATCCTCGGTGTCGAAATCATTAGGATGGCCAAGGCCAAGGCCAAGCATGAGTGAT 2460

2461 GGAGTGTCATGGCATGGCATGCCTTGCCTTGGCTTGGCTTCCAAATCATTCTATTGGAGC 2520

2521 TAATAAGATCGGCCGTCAGCTTTTTAAGACATTGTGTTTTTTGAGCTGTGCACATCATCA 2580

2581 ATTTGCTAACAGTCCGGGTCCTCCATTTTGCATCTTTCTTAGACATTTGCATGACTTTCT 2640

2641 AACTATCATCAGGTGAGCGGCTTTATGGATTTGGTTAGGGCGTCCAATTACTTGTATGTT 2700

2701 GCTTGTTTTGTTATACACTGATTGCGAAACAACTTGTGAGTGAGATTACTTCTAATCCAG 2760

2761 TTTTAGGGGGGCACCTACCTATAGGAAGATGAAACATCATCATTATTTTGGTTGTTTTGC 2820

2821 CTATTACATCCATCTTTTGTACTACTACAATCTGGTTTTTCCATGATGTATGACCTGGTG 2880

2881 GTTACATCAGGTTGATTGGTGCAATGTTAGCATATTCAGTTGTTGTTCATTATAACTTTA 2940

2941 TAATGCCAGATCTTTCCATTTCCAACTGGATTTAGTATTATAAATTGAGTAGTACCCTTT 3000

3001 CTTCATTATTGCTGTCAATATTTGAAGCCTCTAGAAAGAAAGGCTGATAATTGTCAATTC 3060

3061 ATGTCCTTTTGTGTGATCCCACACCAATCAGTGCTTAGATTTAACTTACTCTTCTTTACA 3120

3121 AAAAGGCCTACTTGGATGGCGCTAGAATGATAAGAAAAATTCTTTAAGATGAAACCTTTA 3180

3181 TTTTTGGTATGTCGATTGGAGTTATGAATGTTTAAGCTGAATAAAAAACCCATGACAGGC 3240

3241 ATCCTAGCACAAGAATTTTGTTTGATCCCCTGATCTAACACTACCTGGCTTGTTTTCCAG 3300

3301 GACCCCTTTTCTATTCACACTGGGCAATCAAATATAATTGTTCCTCACATCAATCAGTGC 3360

3361 TTAGATTTGAACATACTCTTAAAGGGGTGCATGGATGGTGCTAGACTGATAAAAGCCTGA 3420

3421 ATCAAATATAATTGTTCCTCACATCAATCAGTGCTTAGATTTGAACATACTCTTAAAGGG 3480

3481 GTGCATGGATGGTGCTAGACTGATAAAAGCAATTCTTGTGGATGCTGCCCGGACTTATGG 3540

3541 ATGTTAAGCCTGAATAAAGAAAGCCATGACAGGCACCGGCATCCCAATCTGGCTTGTTTT 3600

3601 GCACTCATTTTCTATTCACACTGGGCAATCTCGTATAATAGTTGCGTGCAGAACGTCTTT 3660

3661 ATGATCTTTAAATCTTTATACGCATCTCCCTTTTGAACGTGTTGCAATCAAGAGTACTGC 3720

3721 TGCATTCGTCTGATATTCGGGTTGAAACTTTGCAGGTATCCTACCGATGGCGACAAGCAA 3780

387 P T D G D K Q 393

Homeodomain

3781 ATGCTGGCCAAGCAGACTGGCCTAACAAGGAATCAGGTGAACAGCAAAGCTGGTCGCTGT 3840

394 M L A K Q T G L T R N 404

3841 TACCACTAGCATGTCATTTTGTTTGTGGAGCTTGGTTGACGTGTTGGGTTAATAATGGCG 3900

3901 GTGCAGGTGTCGAACTGGTTCATCAACGCGAGGGTGCGGCTCTGGAAGCCAATGGTGGAG 3960

405 Q V S N W F I N A R V R L W K P M V E 423

Homeodomain

3961 GAGATCCACAACCTGGAGATGAGGCAGGTCCACAAGCACTCGCCGCACGACAAGGGCCAG 4020

424 E I H N L E M R Q V H K H S P H D K G Q 443

4021 CAGAACGGCGTCCACGGCCAGGCTCACCAACACTCGTCGCAGCAGCAGCAGCAGCAGCGC 4080

444 Q N G V H G Q A H Q H S S Q Q Q Q Q Q R 463

4081 AGCGGCAAGCGCTCCGAGCCCTGCGACTCGCACCCCGGCCAGAGCAGCGGCGTCACCAGG 4140

464 S G K R S E P C D S H P G Q S S G V T R 483

4141 AACCACCACCACCACAACGCCGCCGCCGCCGCCTCCTCCCACGGTGGTGGCTTCCCGGAC 4200

484 N H H H H N A A A A A S S H G G G F P D 503

4201 GACCTCTCCCAGATGTCCCACTCCATGCAGCAGGGCCAGGTGACCTTCGCCGGGTACGGC 4260

504 D L S Q M S H S M Q Q G Q V T F A G Y G 523

4261 GCGCTGCCCTCCCAGCAGCAGCACGGCAGCATGGCGTCGCCGCAGCACCACCACCACCAT 4320

524 A L P S Q Q Q H G S M A S P Q H H H H H 543

4321 CACGTCGGCGTCGGCGTCGGCGGGGCGGTGAATGCTGGCGGCGGTGGCGGCGTGTCGCTC 4380

544 H V G V G V G G A V N A G G G G G V S L 563

4381 ACCCTCGGCCTCCACCAGAACAACAGGGTCTGCTTCGGGGAGCCGCTGCCGGCCAACCTG 4440

564 T L G L H Q N N R V C F G E P L P A N L 583

4441 GCGCACCGGTTCGGGCTGGAGGACGTCGTCAGCGACCCCTACGTGATGGGCTCGTTCGGC 4500

584 A H R F G L E D V V S D P Y V M G S F G 603

4501 GGCCAGGACCGGCACTTCGCCAAGGAGATCGGCGGCCACCTCCTCCATGATTTCGTCGGG 4560

604 G Q D R H F A K E I G G H L L H D F V G 623

4561 TGATCGACGCCGCCGGTGCTCAGCTCGCTCCCCGCTGATGCACATTGTTGTAATGTACGC 4620

*

4621 ACGCTACGCACTGCTAGTATGCTAGGATGGTATATAGTAAGTCAATCATCACAATCACTT 4680

4681 GGCGGCTTGGCTCGATGATCGGTCGACATGAACACGGTCGATTCCTTTGGAGGGCGGATG 4740

4741 AAACATTGACATGCCCTCTCTCCCATGTACTAGATCCAAAATTCAAACAGCTGTCCAACA 4800

4801 TTGCTCTCAAACACCTTGTGGGCTACACAGAGCTTGAAGAGCAATGCTAGACGTACGGGC 4860

4861 GTTGTACGGGGGATTTACAGGCTCGTTGACTGTGATTGGTTGGAATATGATTAAGGGGGA 4920

4921 CGGCCCCACCCTGAAAATCAGGGGGGAAGCAATTAGATTAGGCCACGGTGTCATCCTGTA 4980

4981 AAACTTTGTACGATCATCCTGTACGTGTAGTATTATTGGAGCTTGAAAATACCTTCTTCC 5040

5041 AAACGGCTTTATGGTCTCCTGCCATTTGTCCTAGTAGAGCTTTTGGATTATCCTTTGGTA 5100

5101 ATTAATGAAATGCACGCATGTGTTCCCACATGGCCAAGGAAAAGTTACCTCTTGTTTATG 5160

5161 CCCAAACGTAAATACCGTGCTGCAGCTCCTTGTTTATGCCCAAAAGTTTTACTACCATTA 5220

5221 CTGTGCCGCAGCTGAAGCTTTCAACCGGTCTCTGTAGAATATGACCATCTTCCGACCTGT 5280

5281 TATGTTAGCCTTTCGATGATATGATATGAACGAAACCTGCTGTATTAGTTAGTCTTTTGA 5340

5341 TGTGCGGTTATATTAATAGTACTAATCACGCCGTCGTTTCCGTTATCCCCTGTATACATC 5400

5401 GATGGGGGTAGCCTCCTAAAGTCTGGACAGCCCATGATTTGGACGCCTGTTGACCTCTAA 5460

5461 TTGGCCCCGTCACGCTAACAAATTATCTATCATCGGGGCTAGCAGCGGTAGATTCGGCCT 5520

5521 ACCCTTTCGATCACGGATCCACGCACTAAACTAATGCCACAGTGCCCCAAATGGCATGGC 5580

5581 ATGGCCATGATGATGGATCCTGGCTGCGCTGTCAGTGTCGTGCTGTGTAGAAGTATTATA 5640

5641 TTATCACTAGTAGGAGTATTATATTAAGCCCATGCATGGTAGAGCATGGTAATAGTACCT 5700

5701 GCCGCTGCATGGTTAACCGTGCTGTTCCTGTCCGGTTTCGCAGCGGTAGTATCGGATCTG 5760

5761 TATATATGTGTCTGGCGCGATGATTCTATAGGTGCGCTGTGTGTGTTGGCTGGTGATTGG 5820

5821 TGATGGGCAGCAGCAGCGTATCGGCCGTGGGTGGGGGTGGCGCCATGCACGTTGTTTTTC 5880

5881 CCGGGAGGGAATCCGCGCGCGCTGTAACTAGGGGGCTAGGGAGTACTTTGGCCTGCAGAG 5940

5941 ACTGCAGTGACTACAGCTCACGTTTTTAAAAGCGTCACTGTTTGCCTCCTCTCTTAGGCG 6000

6001 TGTCAGCTGCTACTGGTGGTGGATGGGGAGCGTAATTGTTCTTTCAATGTGTTGTTGTTG 6060

6061 GTCATGCAATGACCACTGTTACTACACATGGGTGATGGGCCGAGCCTGTCTGAGCTACCA 6120

6121 TAACTCGAAAGATCAGTTAAAAACATAATATCTCACATGGAATCAGAACCATGCAAGTGA 6180

6181 TATAGTACTCCTCTTGTTTACTTATATGTCCTGGCTGCATGGACTCTGGACATCACATCA 6240

6241 GCTAAGCTTGCCATCTCAGCGTAGAGAAAGCCAAGTTTGCTATTGGATGGCCTGTTAATT 6300

6301 TTGTGTGTTGTACTGCTGTTTATTCATGGCCATA 6334

Fig. S1 The structure of TaqSH1-D in Chinese Spring. The translated bases are 215–991, 1870–2249, 3759–3813, and 3904–4560 bp and the deduced polypeptide consists of 623 residues. Untranslated region sequences are double-underlined, and intron sequences are underlined. The stop codon is indicated by an asterisk. A 189-bp insertion in Chinese Spring compared to synthetic wheat S-6214 is indicated by gray shading. The adenine underlined in bold in exon 4 (at bp 4050) is a synonymous SNP in the parental lines (guanine in S-6214). The adenine underlined in bold at bp 5955 is thymine in S-6214. The three conserved protein domains (SKY, BEL, and homeodomain) are indicated by amino acids typed in bold