This document compares the glutenin subunit composition among different classes of Australian and North American wheat. It finds: 1) Each wheat class is blended from various cultivars for specific end uses, and their glutenin compositions reflect these uses. 2) Winter wheat classes generally have stronger glutenin compositions to compensate for lower protein levels. Australian Prime Hard wheat lacks a strong Glu-D1d allele suited for noodles. 3) Western White wheat lacks one high molecular weight glutenin subunit, and contains a unique Glu-B3 allele, suited for cakes and cookies. 4) Further study of gliadin alleles and recombinant lines is needed to fully understand gluten properties and cultivar development

1. Comparison of glutenin subunit composition among
Australian and North American wheat classes
Tatsuya M. Ikeda and Kanenori Takata
池田 達哉、高田 兼則
National Agriculture and Food Research Organization (NARO)
Western Agricultural Research Center, JAPAN
農研機構（NARO）近畿中国四国農業研究中心
日本、広島

2. Classification of HMW-GS alleles
１ 2*
2
7 Glu-A1
8
12 null
a b c
2
6 21
7 7 7 7 20 13 13 14
16 19 15 17
18
Glu-B1
8 8 8 22
9
12
a b c d e f g h i j k
２.2
2 2 3 ４ ２
５
7 Glu-D1
8
12 10 10
12 12 12 12
a b c d e f
Payne & Lawrence 1983

3. Classification of LMW-GS alleles
Glu-A3 a f g f b c d f f g c f
Glu-B3 a j g h b h b j ad g ac m
Glu-D3 a b c c b b a a c b b c
Ikeda et al. IWGS (2008)

4. Identification methods for Glu-3 alleles
and a temporal standard cultivar set
International collaboration has been going on to unify Glu-3 alleles.
Ikeda et al. Int. Wheat Genet Symp O42 (2008)
Ikeda et al. Int. Wheat Conference. (2010)
Liu et al. BMC Plant Biology 10:124 (2010)

5. Effect of glutenin alleles on gluten network
Wild-type Glu-A3d, Glu-B3g
(over-expression = more network)
Glu-D1d Glu-A1c, Glu-A3e
(Extra cysteine = tighter network) (null type = less network)

6. Effect of Glu-1 alleles on gluten strength
Gluten Glu-1 allele
strength
Glu-A1 Glu-B1 Glu-D1
Strong 7OE+8 (al) 5+10 (d)
13+16 (f), 17+18 (i)
1 (a), 2* (b) 7+8 (b/u), 2+12 (a), 3+12 (b)
7+9 (c)
7 (a), 6+8 (d), 20 (e)
Weak null (c) 4+12 (c)
6.1 (bl)
Based on Payne (1987)

7. Effect of Glu-3 alleles on gluten strength
Gluten Glu-3 allele
strength
Glu-A3 Glu-B3 Glu-D3
Strong g
d b, ab
a, b, c d, h, i a, b, c, d, f
f c l?
Weak
e j

8. Glutenin subunit combination and gluten strength
Strong
Glu-B1al/Glu-D1d Strong Extra strong
Glu-D1d
Glu-1
(HMW-GS)
Medium
Glu-D1a
Glu-A1c
Glu-B1a Weak
Weak Strong
Glu-A3e/f Glu-A3e/f Glu-A3d Glu-B3g
Glu-B3j Glu-B3h Glu-B3b
Glu-3 (LMW-GS)

9. Australian & North American wheat classes
Class Protein Usage
APH (Australian Prime Hard) ~13% Yellow alkaline noodle
ASW (Australian Standard White) ~10.5% White-salted noodle
CWRS (1CW)
(Canada Western Red Spring) ~13.5% Best for Pullman-type bread
DNS (Dark Northern Spring) ~14.0% Bread & Yellow alkaline noodle
HRW (Hared Red Winter) ~11.5% Bread & Yellow alkaline noodle
WW (Western White) ~10% Cake & Cookie
Each class blending various cultivars for specific usage.

10. Background & Objectives
• Each wheat classes blended various cultivars for particular usages.
• Studying glutenin compositions of these classes should help us to
understand the relationships with their usages.
• But, commercial cultivars are not available through seed banks.
Materials & Methods
• 60 seeds of wheat classes imported into Japan in 2009.
• Isolating DNA and protein from individual seeds.
• Identifying glutenin alleles by PCR markers, SDS-PAGE and 2D
electrophoresis.

11. SDS-PAGE analysis of glutenin fractions
of wheat classes
DNS HRW ASW(N)
HMW-GS
LMW-GS
& gliadins
SDS-PAGE condition according to Dr.R.J.Peña

12. Glu-3 allele identification
pH6 IEF pH10 pH6 IEF pH10
kDa
3 11
50 3 3 3
12 12
4 1
5 1 5
11 2 3
37 2 4
10 10 8/9
8/9
DNS HRW
Glu-A3d, Glu-B3h, Glu-D3a Glu-A3b, Glu-B3b, Glu-D3c
Combination of PCR, SDS-PAGE, 2DE and N-terminal sequencing.
Ikeda et al. (2006)

13. Observed number of allelic combination
35
30
25
20
15
10
5
0
APH ASW ASWN 1CW DNS HRW WW
= minimum number of cultivars consisting of each class

14. Glutenin subunit composition
Glu-A1 Glu-B1
100%
75%
50%
25%
0%
1CW DNS HRW PH ASW WW
a b c new

15. Glutenin subunit composition
Glu-D1 Glu-A3

16. Glu-3 subunit composition
Glu-B3 Glu-D3

17. Identification of a new Glu-A1 alleles in ASWN
a a
b new new b new b
The new Glu-A1 allele was found in an ASWN cultivar, Calingiri.

18. Identification of a new Glu-3 alleles in HRW
pH6 IEF pH10
3 3
d 4 3
d*
f
12 12 1
5
2 11
10
8/9
Glu-A3d*, Glu-B3ab, Glu-D3f
Glu-A3d* was not found before.
Glu-D3f was found in Cheyenne (Ikeda et al. 2006)

19. Identification of a new Glu-3 alleles in WW
pH6 IEF pH10
12
3
5 1
2 4
10
8/9
Glu-A3c, Glu-B3ae, Glu-D3c
A new Glu-B3 allele (Glu-B3ae） was found among club wheats and
expresses less amount of LMW-GS

20. Glutenin subunit combination and gluten strength
Strong
Glu-B1al/Glu-D1d Strong Extra strong
Glu-D1d
Glu-1
(HMW-GS)
Medium
Glu-D1a
Glu-A1c
Glu-B1a Weak
Weak Strong
Glu-A3e Glu-A3e/f Glu-A3d Glu-B3g
Glu-B3j Glu-B3h Glu-B3b
Glu-3 (LMW-GS)

21. Glutenin subunit combination and gluten strength
Strong
Glu-B1al/Glu-D1d Strong Extra strong
DNS
1CW
Glu-D1d HRW
ASW
Glu-1
APH
(HMW-GS)
Medium
Glu-D1a
Weak
Glu-A1c Club WW
Glu-B1a
Weak Strong
Glu-A3e Glu-A3e/f Glu-A3d Glu-B3g
Glu-B3j Glu-B3h Glu-B3b
Glu-3 (LMW-GS)

22. Wheat breeding targets
Strong
Glu-B1al/Glu-D1d Blending
Pullman-type bread（11.5～13.5%） （>12％）
Glu-D1d
Yellow alkaline
noodle
Glu-1
Glu-D1a Noodle European
（8.0～12.5%） bread
Cake & Cookies
（<10%）
Glu-A1c
Glu-B1a
Strong
Weak Glu-A3e Glu-B3h Glu-A3d Glu-B3g
Glu-B3j Glu-B3b
Glu-3

23. Results & Discussion
• Glutenin compositions of each class appear to reflect end-use properties.
• Most of winter wheat classes (HRW, ASW) have glutenin composition
expected to increase gluten strength, which might compensate relatively
lower protein content.
• Majority of APH lacked Glu-D1d allele. It might suitable for the texture of
yellow alkaline noodle.
• Most of WW lacked one (not two) of HMW-GSs. Club wheat also
contains a unique Glu-B3 allele (Glu-B3ae). These compositions might
be suitable for cake and cookie making.
• It is necessary to clarify the effect of gliadin allele compositions on
gluten properties (extensibility). We need recombinant lines breaking
Glu-3/Gli-1 linkages (e.g. Glu-B3h/Gli-B1d).

24. Further international collaboration
for gluten research
Proposal of International Gluten Research Group as a
part of IRIWI (International Research Initiative for
Wheat Improvement)
Activities Exchanging and sharing valuable wheat cultivars.
Unifying the nomenclature of gluten protein alleles and the
standard methods to examine gluten proteins.
Studying the usefulness of particular gluten compositions for
quality under various environmental conditions.
Goals
Development of wheat cultivars with specific grain quality
attributes under different environmental conditions, including
heat and drought stresses.

25. We need further collaboration
Thank you!