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

Classification of HMW-GS alleles

Glu-B1

Payne & Lawrence 1983

12

2

7

8

12

2

7

8

12

2

7

8

12

2 3 ４ ５ ２

２.2

12 12 10 10

12

7 7 7

8 8 9

6

20 13 16 19

13 14 15 17

18

21

22

１ 2*

a b c

a b c d e f g h i j k

a b c d e f

Glu-A1

Glu-D1

null

Classification of LMW-GS alleles

a f g f b c d f f c f g

a j g h b h b j ad ac m g

a b c c b b a a c b c b

Glu-A3

Glu-B3

Glu-D3

Ikeda et al. IWGS (2008)

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)

Identification methods for Glu-3 alleles

and a temporal standard cultivar set

Glu-D1d

(Extra cysteine = tighter network)

Glu-A1c, Glu-A3e

(null type = less network)

Glu-A3d, Glu-B3g

(over-expression = more network)

Effect of glutenin alleles on gluten network

Wild-type

Effect of Glu-1 alleles on gluten strength

Gluten

strength Glu-1 allele

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)

Weak null (c) 7 (a), 6+8 (d), 20 (e)

6.1 (bl) 4+12 (c)

Based on Payne (1987)

Gluten

strength Glu-3 allele

Glu-A3 Glu-B3 Glu-D3

Strong g

d b, ab

a, b, c d, h, i a, b, c, d, f

Weak f c l?

e j

Effect of Glu-3 alleles on gluten strength

Glutenin subunit combination and gluten strength

Strong

Weak Strong

Glu-3 (LMW-GS)

Glu-1

(HMW-GS)

Glu-A3d

Glu-B3b

Glu-D1d

Glu-B3g Glu-A3e/f

Glu-B3h Glu-A3e/f

Glu-B3j

Glu-D1a

Glu-A1c

Glu-B1a

Extra strong Strong

Medium

Weak

Glu-B1al/Glu-D1d

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.

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.

DNS HRW

SDS-PAGE analysis of glutenin fractions

of wheat classes

HMW-GS

LMW-GS

& gliadins

ASW(N)

SDS-PAGE condition according to Dr.R.J.Peña

H 6 D a

3 7

5 0

p k

IEF pH10

3 3

8/9 10

4

2 1 5

11 3

3

8/9 10

4

1 5

11

H 6 p IEF pH10

HRW DNS

Glu-A3b, Glu-B3b, Glu-D3c Glu-A3d, Glu-B3h, Glu-D3a

Combination of PCR, SDS-PAGE, 2DE and N-terminal sequencing.

2

Glu-3 allele identification

3

12 12

Ikeda et al. (2006)

Observed number of allelic combination

= minimum number of cultivars consisting of each class

0

5

10

15

20

25

30

35

APH ASW ASWN 1CW DNS HRW WW

Glutenin subunit composition

Glu-A1 Glu-B1

0%

25%

50%

75%

100%

1CW DNS HRW PH ASW WWa b c new

Glutenin subunit composition

Glu-D1 Glu-A3

Glu-3 subunit composition

Glu-B3 Glu-D3

The new Glu-A1 allele was found in an ASWN cultivar, Calingiri.

Identification of a new Glu-A1 alleles in ASWN

a b b

a b new new new

H 6 p IEF pH10

Glu-A3d* was not found before.

Glu-D3f was found in Cheyenne (Ikeda et al. 2006)

3 3

8/9 10

4

2 1 5

11

Glu-A3d*, Glu-B3ab, Glu-D3f

3

12 12

Identification of a new Glu-3 alleles in HRW

d d*

f

H 6 p IEF pH10

A new Glu-B3 allele (Glu-B3ae） was found among club wheats and

expresses less amount of LMW-GS

3

8/9

10

4 2 1 5

12

Glu-A3c, Glu-B3ae, Glu-D3c

Identification of a new Glu-3 alleles in WW

Glutenin subunit combination and gluten strength

Strong

Weak Strong

Glu-3 (LMW-GS)

Glu-1

(HMW-GS)

Glu-A3d

Glu-B3b

Glu-D1d

Glu-B3g Glu-A3e

Glu-B3j

Glu-D1a

Glu-A1c

Glu-B1a

Extra strong Strong

Medium

Weak

Glu-B1al/Glu-D1d

Glu-A3e/f

Glu-B3h

Glutenin subunit combination and gluten strength

Strong

Weak Strong

Glu-3 (LMW-GS)

Glu-1

(HMW-GS)

Glu-A3d

Glu-B3b

Glu-D1d

Glu-B3g Glu-A3e

Glu-B3j

Glu-D1a

Glu-A1c

Glu-B1a

Extra strong Strong

Medium

Weak

Glu-B1al/Glu-D1d DNS

1CW HRW

WW

ASW

APH

Club

Glu-A3e/f

Glu-B3h

Strong

Weak Strong

Blending

（>12％） Pullman-type bread（11.5～13.5%）

Noodle （8.0～12.5%）

Cake & Cookies （<10%）

Glu-3

Glu-1

Wheat breeding targets

Yellow alkaline

noodle

European

bread

Glu-D1d

Glu-D1a

Glu-A1c

Glu-B1a

Glu-B1al/Glu-D1d

Glu-A3d

Glu-B3b

Glu-B3g Glu-B3h Glu-A3e

Glu-B3j

• 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).

Results & Discussion

Further international collaboration

for gluten research

Proposal of International Gluten Research Group as a

part of IRIWI (International Research Initiative for

Wheat Improvement)

Unifying the nomenclature of gluten protein alleles and the

standard methods to examine gluten proteins.

Exchanging and sharing valuable wheat cultivars.

Studying the usefulness of particular gluten compositions for

quality under various environmental conditions.

Development of wheat cultivars with specific grain quality

attributes under different environmental conditions, including

heat and drought stresses.

Activities

Goals

We need further collaboration

Thank you!