Table S1. Pedigree of selected wheat D-genome synthetic hexaploids used in this study.
S. No / Name / Pedigree / S.No / Name / Pedigree1 / SH - 6 / DOY1 / AE.SQUARROSA (188)* / 24 / SH - 398 / DOY1 / AE.SQUARROSA (334)
2 / SH - 43 / DOY1 / AE.SQUARROSA (216) / 25 / SH - 401 / DOY1 / AE.SQUARROSA (372)
3 / SH - 115 / DOY1 / AE.SQUARROSA (446) / 26 / SH - 409 / DOY1 / AE.SQUARROSA (418)
4 / SH - 117 / DOY1 / AE.SQUARROSA (447) / 27 / SH - 422 / DOY1 / AE.SQUARROSA (516)
5 / SH - 123 / DOY1 / AE.SQUARROSA (488) / 28 / SH - 424 / DOY1 / AE.SQUARROSA (517)
6 / SH - 128 / DOY1 / AE.SQUARROSA (510) / 29 / SH - 426 / DOY1 / AE.SQUARROSA (526)
7 / SH - 129 / DOY1 / AE.SQUARROSA (511) / 30 / SH - 434 / DOY1 / AE.SQUARROSA (1016)
8 / SH - 131 / DOY1 / AE.SQUARROSA (515) / 31 / SH - 441 / DOY1 / AE.SQUARROSA (1024)
9 / SH - 302 / DOY1 / AE.SQUARROSA (293) / 32 / SH - 444 / DOY1 / AE.SQUARROSA (1027)
10 / SH - 323 / DOY1 / AE.SQUARROSA (333) / 33 / SH - 447 / DOY1 / AE.SQUARROSA (1030)
11 / SH - 326 / DOY1 / AE.SQUARROSA (349) / 34 / SH - 500 / DOY1 / AE.SQUARROSA (1018)
12 / SH - 330 / DOY1 / AE.SQUARROSA (370) / 35 / SH - 505 / DOY1 / AE.SQUARROSA (1026)
13 / SH - 341 / DOY1 / AE.SQUARROSA (428) / 36 / SH - 509 / DOY1 / AE.SQUARROSA (1029)
14 / SH - 349 / DOY1 / AE.SQUARROSA (458) / 37 / SH - 618 / DOY1 / AE.SQUARROSA (172)
15 / SH - 361 / DOY1 / AE.SQUARROSA (507) / 38 / SH - 638 / DOY1 / AE.SQUARROSA (295)
16 / SH - 363 / DOY1 / AE.SQUARROSA (532) / 39 / SH - 648 / DOY1 / AE.SQUARROSA (359)
17 / SH - 373 / DOY1 / AE.SQUARROSA (177) / 40 / SH - 649 / DOY1 / AE.SQUARROSA (360)
18 / SH - 377 / DOY1 / AE.SQUARROSA (255) / 41 / SH - 672 / DOY1 / AE.SQUARROSA (518)
19 / SH - 380 / DOY1 / AE.SQUARROSA (256) / 42 / SH - 675 / DOY1 / AE.SQUARROSA (534)
20 / SH - 381 / DOY1 / AE.SQUARROSA (258) / 43 / SH - 677 / DOY1 / AE.SQUARROSA (540)
21 / SH - 385 / DOY1 / AE.SQUARROSA (267) / 44 / SH - 678 / DOY1 / AE.SQUARROSA (1010)
22 / SH - 395 / DOY1 / AE.SQUARROSA (318) / 45 / SH - 852 / DOY1 / AE.SQUARROSA (632)
23 / SH - 396 / DOY1 / AE.SQUARROSA (322)
*Ae. tauschii accession numbers maintained at CIMMY gene bank
Table S2. Details of 10 SSR primers used for genetic diversity analysis in SHWs
S.No / Primer name / Forward & reverse primer / Repeat motifs / Annealing / Locustemperature
1 / Xgwm295 / 5’ GTGAAGCAGACCCACAACAC3’ / (GA)25 / 60 / 7D
5'GACGGCTGCGACGTAGAG3’
2 / Xbarc76 / 5’ATTCGTTGCTGCCACTTGCTG3’ / (TATC)7 / 58 / 7D
5’GCGCGACACGGAGTAAGGACACC3’
3 / Xbarc126 / 5’CCATTGAAACCGGATTTGAGTCG3’ / (CTT)17 / 52 / 7D
5’CGTTCCATCCGAAATCAGCAC3’
4 / Xbarc214 / 5’CGCTTTCGGGACAGTGAAGGTGTAT3’ / (CT)16 / 52 / 7D
5’CGGTACGCGCGAGGAGGAAGAAGG3’
5 / Xbarc53 / 5’GCGTCGTTCCTTTGCTTGTACCAGTA3’ / (TAGA)7 / 58 / 7D
5’GCGCGTCCTTCCAATGCAGAGTAGA3’
6 / Cfd41 / 5’TAAAGTCTCAGGCGACCCAC3’ / (CA)17 / 60 / 7D
5’AGTGATAGACGGATGGCACC3’
7 / Cfd14 / 5’CCACCGGCCAGAGTAGTATT3’ / (TC)28 / 60 / 7D
5’TCCTGGTCTAACAACGAGAAGA3’
8 / Cfd46 / 5’TGGTGGTATAGTCGTTGGAGC3’ / (GT)29 / 60 / 7D
5’CCACACACACACACCATCAA3’
9 / Wmc405 / 5’GTGCGGAAAGAGACGAGGTT3’ / (CA)21 / 61 / 7D
5’TATGTCCACGTTGGCAGAGG3’
10 / Wmc121 / 5’GGCTGTGGTCTCCCGATCATTC3’ / (GA)12 / 61 / 7D
5’ACTGGACTTGAGGAGGCTGGCA3’
Table S3. Basic statistics for root and shoot length response of synthetic hexaploids to different boron treatments
Character / Control / 0.01M / 0.025MShoot Length / Min / 2.77 / 0.51 / 0.07
Max / 18.76 / 9.54 / 4.64
Mean / 9.26 / 5.14 / 2.10
St Dev. / 4.06 / 2.15 / 1.10
St. Error / 0.60 / 0.32 / 0.16
Heritability / 0.65 / 0.74 / 0.82
Root Length / Min / 3.30 / 1.48 / 0.02
Max / 17.49 / 6.35 / 2.89
Mean / 8.66 / 4.32 / 1.21
St Dev. / 3.98 / 1.29 / 0.84
St. Error / 0.59 / 0.19 / 0.13
Heritability / 0.80 / 0.85 / 0.88
Table S4. Analysis of variance (ANOVA) for synthetic hexaploids in different boron treatments, (a) root length and (b) shoot length at different boron treatments
a)
SOV / df / SS / MS / F / PMain Effect
Variety / 44 / 796.40 / 18.10 / 8.10 / 0.0000***
Treatment / 2 / 2444.35 / 1222.17 / 547.11 / 0.0000***
Variety*Treatment / 88 / 929.03 / 10.55 / 4.72 / 0.000***
LSD (Varieties) = 2.25; LSD (Treatments) = 0.58; CV (%) =31.85
b)
SOV / df / SS / MS / F / pMain Effects
Variety / 44 / 1093.78 / 24.86 / 4.56 / 0.000***
Treatment / 2 / 2160.42 / 1080.21 / 198.26 / 0.000***
Variety*Treatment / 88 / 1146.75 / 13.03 / 2.39 / 0.000***
LSD (Varieties) = 3.52; LSD (Treatments) = 0.90; CV (%) =43.10
Table S5. Root length and relative root length response in SHWs at different boron treatments
Root length (cm) / RRL (%)Genotypes / R / Control / 0.01M / 0.025M / Mean / RGS (%) / 0.01M / 0.025M
SH-505 / T / 7.01 / 5.99 / 1.99 / 5.00 / 43.08 / 85.45 / 28.39
SH-380 / T / 5.84 / 5.49 / 1.26 / 4.20 / 42.21 / 94.01 / 21.58
SH-381 / T / 4.68 / 3.78 / 1.42 / 3.29 / 44.44 / 80.77 / 30.34
SH-117 / T / 4.33 / 5.09 / 0.58 / 3.33 / 34.64 / 117.55 / 13.39
SH-302 / T / 2.44 / 2.61 / 0.02 / 1.69 / 46.11 / 106.97 / 0.82
SH-131 / T / 3.30 / 4.61 / 0.96 / 2.96 / 15.61 / 139.70 / 29.09
SH-361 / T / 2.14 / 2.19 / 0.50 / 1.61 / 37.15 / 102.34 / 23.36
SH-363 / T / 3.36 / 2.88 / 0.86 / 2.37 / 44.35 / 85.71 / 25.60
SH-618 / T / 3.40 / 3.35 / 2.29 / 3.01 / 17.06 / 98.53 / 67.35
SH-649 / T / 5.41 / 4.47 / 2.39 / 4.09 / 36.60 / 82.62 / 44.18
SH-852 / T / 5.06 / 4.05 / 1.34 / 3.48 / 46.74 / 80.04 / 26.48
SH-677 / MT / 5.05 / 3.48 / 1.23 / 3.25 / 53.37 / 68.91 / 24.36
SH-341 / MT / 7.11 / 5.67 / 0.82 / 4.53 / 54.36 / 79.75 / 11.53
SH- 6 / MT / 6.65 / 5.72 / 0.20 / 4.19 / 55.49 / 86.02 / 3.01
SH-323 / MT / 5.76 / 4.30 / 1.06 / 3.71 / 53.47 / 74.65 / 18.40
SH-441 / MT / 7.00 / 3.91 / 2.31 / 4.41 / 55.57 / 55.86 / 33.00
SH-444 / MT / 8.09 / 5.13 / 1.83 / 5.02 / 56.98 / 63.41 / 22.62
SH-509 / MT / 9.73 / 6.35 / 2.85 / 6.31 / 52.72 / 65.26 / 29.29
SH-123 / MS / 6.91 / 4.44 / 0.37 / 3.90 / 65.20 / 0.64 / 0.05
SH-326 / MS / 8.84 / 4.57 / 2.28 / 5.23 / 61.26 / 51.70 / 25.79
SH-129 / MS / 3.92 / 2.79 / 0.21 / 2.31 / 61.73 / 71.17 / 5.36
SH-648 / MS / 11.52 / 6.21 / 1.78 / 6.50 / 65.32 / 53.91 / 15.45
SH-675 / MS / 12.24 / 4.68 / 2.55 / 6.49 / 70.47 / 38.24 / 20.83
SH-43 / S / 5.67 / 2.54 / 0.08 / 2.77 / 76.71 / 0.45 / 0.014
SH-115 / S / 11.15 / 4.91 / 0.48 / 5.51 / 75.83 / 44.04 / 4.30
SH-128 / S / 9.73 / 5.00 / 0.74 / 5.16 / 70.50 / 51.39 / 7.61
SH-330 / S / 14.20 / 3.83 / 1.00 / 6.34 / 82.99 / 26.97 / 7.04
SH-349 / S / 14.61 / 5.18 / 1.31 / 7.03 / 77.79 / 35.46 / 8.97
SH-373 / S / 4.94 / 2.03 / 0.18 / 2.38 / 77.63 / 41.09 / 3.64
SH-377 / S / 13.68 / 6.22 / 1.83 / 7.24 / 70.58 / 45.47 / 13.38
SH-385 / S / 7.58 / 2.75 / 0.02 / 3.45 / 81.73 / 36.28 / 0.26
SH-395 / S / 15.60 / 5.70 / 1.79 / 7.70 / 75.99 / 36.54 / 11.47
SH-396 / S / 14.16 / 6.15 / 1.01 / 7.11 / 74.72 / 43.43 / 7.13
SH-398 / S / 11.69 / 5.70 / 1.04 / 6.14 / 71.17 / 48.76 / 8.90
SH-401 / S / 12.57 / 3.97 / 0.17 / 5.57 / 83.53 / 31.58 / 1.35
SH-409 / S / 12.49 / 4.88 / 1.96 / 6.44 / 72.62 / 39.07 / 15.69
SH-422 / S / 10.83 / 4.40 / 0.11 / 5.11 / 79.18 / 40.63 / 1.02
SH-424 / S / 11.21 / 3.76 / 0.22 / 5.06 / 82.25 / 33.54 / 1.96
SH-426 / S / 17.49 / 6.32 / 1.91 / 8.57 / 76.47 / 36.13 / 10.92
SH-434 / S / 15.08 / 4.86 / 0.91 / 6.95 / 80.87 / 32.23 / 6.03
SH-447 / S / 8.05 / 1.48 / 0.54 / 3.36 / 87.45 / 18.39 / 6.71
SH-500 / S / 7.93 / 3.13 / 1.29 / 4.12 / 72.13 / 39.47 / 16.27
SH-638 / S / 8.86 / 2.76 / 2.55 / 4.72 / 70.03 / 31.15 / 28.78
SH-672 / S / 10.90 / 3.39 / 2.89 / 5.73 / 71.19 / 31.10 / 26.51
SH-678 / S / 11.52 / 3.56 / 1.27 / 5.45 / 79.04 / 30.90 / 11.02
RGS: Root growth suppression, RRL: Relative root length, R: Response to different boron treatment.
Table S6. Shoot length and relative shoot length response of SHWs at different boron treatments.
Shoot length (cm) / RSL (%)Genotypes / R / Control / 0.01M / 0.025M / Mean / SGS (%) / BO1 / BO2
SH- 6 / T / 6.16 / 9.54 / 0.83 / 5.51 / 15.83 / 154.87 / 13.47
SH-117 / T / 4.08 / 6.40 / 1.61 / 4.03 / 1.84 / 156.86 / 39.46
SH-123 / T / 6.50 / 7.48 / 1.42 / 5.13 / 31.54 / 115.08 / 21.85
SH-129 / T / 3.97 / 4.63 / 1.14 / 3.25 / 27.33 / 116.62 / 28.72
SH-131 / T / 6.10 / 8.20 / 2.01 / 5.44 / 16.31 / 134.43 / 32.95
SH-323 / T / 5.50 / 7.38 / 1.66 / 4.85 / 17.82 / 134.18 / 30.18
SH-441 / T / 6.15 / 5.16 / 2.80 / 4.70 / 35.28 / 83.90 / 45.53
SH-505 / T / 9.01 / 5.92 / 3.59 / 6.17 / 47.23 / 65.70 / 39.84
SH-618 / T / 3.36 / 2.29 / 2.60 / 2.75 / 27.23 / 68.15 / 77.38
SH-361 / T / 2.77 / 3.75 / 0.81 / 2.44 / 17.69 / 135.38 / 29.24
SH-380 / T / 5.73 / 6.29 / 2.96 / 4.99 / 19.28 / 109.77 / 51.66
SH-677 / T / 3.53 / 3.25 / 2.16 / 2.98 / 23.37 / 92.07 / 61.19
SH-43 / MT / 4.75 / 4.02 / 0.52 / 3.10 / 52.21 / 84.63 / 10.95
SH-302 / MT / 2.92 / 2.26 / 0.07 / 1.75 / 60.10 / 77.40 / 2.40
SH-377 / MT / 9.39 / 6.52 / 2.28 / 6.06 / 53.14 / 69.44 / 24.28
SH-381 / MT / 5.32 / 3.33 / 1.44 / 3.36 / 55.17 / 62.59 / 27.07
SH-396 / MT / 13.08 / 8.02 / 3.02 / 8.04 / 57.80 / 61.31 / 23.09
SH-426 / MT / 12.92 / 7.16 / 3.33 / 7.80 / 59.40 / 55.42 / 25.77
SH-852 / MT / 4.59 / 1.93 / 2.38 / 2.97 / 53.05 / 42.05 / 51.85
SH-398 / MT / 9.58 / 6.70 / 2.45 / 6.24 / 52.24 / 69.94 / 25.57
SH-648 / MT / 12.67 / 7.01 / 3.06 / 7.58 / 60.26 / 55.33 / 24.15
SH-649 / MT / 7.76 / 3.84 / 3.48 / 5.03 / 52.84 / 49.48 / 44.85
SH-672 / MT / 10.98 / 6.55 / 3.84 / 7.12 / 52.69 / 59.65 / 34.97
SH-115 / MS / 14.91 / 8.15 / 1.29 / 8.12 / 68.34 / 54.66 / 8.65
SH-128 / MS / 11.96 / 7.22 / 1.53 / 6.90 / 63.42 / 60.37 / 12.79
SH-349 / MS / 12.79 / 5.49 / 2.23 / 6.84 / 69.82 / 42.92 / 17.44
SH-363 / MS / 4.00 / 1.42 / 1.42 / 2.28 / 64.50 / 35.50 / 35.50
SH-395 / MS / 11.80 / 5.80 / 2.12 / 6.57 / 66.44 / 49.15 / 17.97
SH-401 / MS / 10.91 / 5.90 / 1.12 / 5.98 / 67.83 / 54.08 / 10.27
SH-409 / MS / 13.04 / 5.82 / 2.68 / 7.18 / 67.41 / 44.63 / 20.55
SH-422 / MS / 11.25 / 7.05 / 1.14 / 6.48 / 63.60 / 62.67 / 10.13
SH-500 / MS / 10.88 / 2.81 / 3.74 / 5.81 / 69.90 / 25.83 / 34.38
SH-509 / MS / 16.26 / 7.04 / 4.41 / 9.24 / 64.79 / 43.30 / 27.12
SH-675 / MS / 18.76 / 7.43 / 4.64 / 10.28 / 67.83 / 39.61 / 24.73
SH-326 / S / 11.12 / 4.32 / 1.63 / 5.69 / 73.25 / 38.85 / 14.66
SH-330 / S / 11.05 / 4.42 / 1.67 / 5.71 / 72.44 / 40.00 / 15.11
SH-341 / S / 10.36 / 4.58 / 1.21 / 5.38 / 72.06 / 44.21 / 11.68
SH-373 / S / 6.32 / 2.55 / 0.99 / 3.29 / 71.99 / 40.35 / 15.66
SH-385 / S / 8.27 / 0.51 / 0.36 / 3.05 / 94.74 / 6.17 / 4.35
SH-424 / S / 12.31 / 3.10 / 0.36 / 5.26 / 85.95 / 25.18 / 2.92
SH-434 / S / 16.00 / 6.68 / 2.06 / 8.25 / 72.69 / 41.75 / 12.88
SH-444 / S / 12.69 / 4.46 / 2.79 / 6.65 / 71.43 / 35.15 / 21.99
SH-447 / S / 12.26 / 1.78 / 1.74 / 5.26 / 85.64 / 14.52 / 14.19
SH-638 / S / 12.43 / 3.56 / 3.06 / 6.35 / 73.37 / 28.64 / 24.62
SH-678 / S / 10.57 / 3.40 / 2.68 / 5.55 / 71.24 / 32.17 / 25.35
SGS: Shoot growth suppression, RSL: Relative shoot length, R: Response to different boron treatment
Table S7. Boron toxicity symptoms in synthetic hexaploids at adult plant stage
Damage (%) / Damage (%)S.No / Genotypes / 0 mg/kg / 25 mg/kg / Symptoms / 50 mg/kg / damage / Mean
1 / SH-6 / 0 / 10 / Initial / 10 / Initial / 10
2 / SH-43 / 0 / 55 / Very heavy / 55 / Very heavy / 55
3 / SH-115 / 0 / 50 / Heavy / 40 / heavy / 45
4 / SH-117 / 0 / 30 / Moderate / 85 / Very heavy / 57.50
5 / SH-123 / 0 / 30 / Moderate / 75 / Very heavy / 52.50
6 / SH-128 / 0 / 35 / Moderate / 55 / Very heavy / 45
7 / SH-129 / 0 / 50 / Heavy / 50 / Heavy / 50
8 / SH-131 / 0 / 35 / Moderate / 80 / Very heavy / 57.50
9 / SH-302 / 0 / 35 / Moderate / 65 / Very heavy / 50
10 / SH-323 / 0 / 13 / Moderate / 75 / Very heavy / 44
11 / SH-326 / 0 / 7 / Initial / 55 / Very heavy / 31
12 / SH-330 / 0 / 25 / Moderate / 85 / Very heavy / 55
13 / SH-341 / 0 / 30 / Moderate / 35 / Moderate / 35
14 / SH-349 / 0 / 25 / Moderate / 55 / Very heavy / 40
15 / SH-361 / 0 / 25 / Moderate / 27 / Moderate / 27
16 / SH-363 / 0 / 10 / Initial / 35 / Moderate / 22.50
17 / SH-373 / 0 / 10 / Initial / 55 / Very heavy / 32.50
18 / SH-377 / 0 / 7 / Initial / 35 / Moderate / 21
19 / SH-380 / 0 / 10 / Initial / 50 / Heavy / 30
20 / SH-381 / 0 / 35 / Moderate / 85 / Very heavy / 60
21 / SH-385 / 0 / 45 / Heavy / 40 / Heavy / 42.50
22 / SH-395 / 0 / 35 / Moderate / 85 / Very heavy / 60
23 / SH-396 / 0 / 7 / Initial / 40 / Heavy / 23.50
24 / SH-398 / 0 / 35 / Moderate / 85 / Very heavy / 60
25 / SH-401 / 0 / 7 / Initial / 50 / Heavy / 28.50
26 / SH-409 / 0 / 35 / Moderate / 85 / Very heavy / 60
27 / SH-422 / 0 / 23 / Moderate / 65 / Very heavy / 65
28 / SH-424 / 0 / 13 / Initial / 60 / Very heavy / 36.50
29 / SH-426 / 0 / 30 / Moderate / 65 / Very heavy / 65
30 / SH-434 / 0 / 45 / Heavy / 85 / Very heavy / 65
31 / SH-441 / 0 / 35 / Moderate / 65 / Very heavy / 50
32 / SH-444 / 0 / 10 / Initial / 25 / Moderate / 25
33 / SH-447 / 0 / 5 / Initial / 25 / Moderate / 25
34 / SH-500 / 0 / 7 / Initial / 10 / Initial / 7
35 / SH-505 / 0 / 7 / Initial / 40 / heavy / 23.50
36 / SH-509 / 0 / 55 / Heavy / 85 / Very heavy / 85
37 / SH-618 / 0 / 25 / Moderate / 35 / Moderate / 30
38 / SH-638 / 0 / 35 / Moderate / 30 / Moderate / 32.50
39 / SH-648 / 0 / 65 / Very heavy / 45 / heavy / 55
40 / SH-649 / 0 / 13 / Moderate / 13 / Moderate / 13
41 / SH-672 / 0 / 65 / Very heavy / 55 / Very heavy / 60
42 / SH-675 / 0 / 13 / Moderate / 45 / heavy / 29
43 / SH-677 / 0 / 65 / Very heavy / 65 / Very heavy / 65
44 / SH-678 / 0 / 45 / Heavy / 65 / Very heavy / 55
45 / SH-852 / 0 / 25 / Moderate / 40 / Moderate / 32.50
Table. 8 showing Major allele frequency, allele No, gene diversity and PIC value for 10 SSR primers.