Supplementary Figure Legends
S.Fig. 1: AS-PCR products of the complete coding region of 1By18 gene from bread wheat
cultivar CB037C. M: molecular mass markers. The amplification product of 1By18 gene
from CB037C marked with an arrow.
S.Fig. 2: Comparison of the deduced amino acid sequence of HMW-GS 1By18 gene with
seven other Glu-1B genes. HMW-GS genes and their GenBank numbers are 1By18
(KF430469), 1By8 (AY245497), 1By9 (X61026), 1B20y (DQ086215), 1By15 (KF733215),
1By16 (EF540765), 1Ay (FJ404595) and 1Dy10 (X12929). The primary structures of HMWGS including signal peptide, N-terminal domain, repetitive domain, and C-terminal domain
are indicated. Seven conserved cysteine residues are highlighted by red shading. Deletions
are indicated by dashes and SNPs are marked by boxes¬¬.
S.Fig. 3: Identification of the heterologously expressed 1By18 protein in E. coli by SDSPAGE (a) and Western blot analysis (b). M: molecular weight markers; 1. pET-28a vector in
BL21; 2. the induced protein of the 1By18 gene; 3. Western blott analysis of the induced
protein. HMW-GS from CB037C and CS were used as the native protein control. The
expressed protein bands are marked with arrows.
S.Fig. 4: Validation of the SNP-based AS-PCR markers (TF1/TR1 and TF1/TR2) for
discrimination of the 1By18 gene using different wheat cultivars. a. Amplification by
TF1/TR1: the target band with 284 bp is indicated. b. Amplification by TF1/TR2: the target
band with 365 bp is indicated. M: DNA size standard. Variety numbers in the figure are same
as in Table 1. M: DNA size standard.
S.Fig. 5: Validation of the SNP-based AS-PCR markers (TF1/TR1 and TF1/TR2) for 1By18
gene using five RILs derived from a CB037C × CS-1Sl/1B cross. a. HMW-GS identification
in five RILs by SDS-PAGE. b. Amplification by TF1/TR1: the target band with 284 bp is
indicated. Line 1, 4: RIL-3; Line 2-3, 5-6: RIL-4; Line 7, 9, 10: RIL-5; Line 8: CB037C; Line
11-13: RIL-1; Line 14-15: RIL-2; Line 16-17: CS-1Sl/1B. c. Amplification by TF1/TR2: the
target band with 365 bp is indicated. Line 18: CS-1Sl/1B; Line 19-20: RIL-3; Line 21: RIL-1;
Line 22: RIL-4; Line 23, 25: RIL-5; Line 24: CB037C; Line 26: RIL-2. M: DNA size
standard.
S.Fig. 1
SFig 2
a
S.Fig 3
b
1 2 3 4 5 7 8 9 25 26 30 31 35 36 39 46 48 69 70 77 78 90 96 104 M
50
500bp
0b
300bp
p
50
0b
500bp
p
25 26 30 31 10 35 36 39 11 46 48 69 70 12 77 78 90 96 M
500bp
50
400bp
0b
p
300bp
S.Fig 4
CS-1Sl(1B) RIL-1
RIL-2
2.3*
2.3*
2.3*
RIL-3
RIL-4
RIL-5
1
2
2
5
2
16*
5
1
5
16*
17
17
17
17
18
18
12
18
12
18
10
10
10
1
2
HMW-GS
16*
a
12
M
1
2
3
4
5
12
6
7
8
9
CB037C
10 11 12 13 14 15 16 17
284bp
300bp
b
M
18
19
20
21
500bp
c
S.Fig 5
22
23
24
25
26