Tatsuya M. Ikeda, W. John Rogers, Gerard Branlard,

Roberto J. Peña, Silvia E. Lerner, Adriana Arrigoni,

Wujun Ma, Rudi Appels, Odean Lukow, William

Hurkman, Marie Appelbee, Mike Sissons, Jose M.

Carrillo and Zhonghu He

11th

IGW

1National Agriculture and Food Research Organization, Hiroshima, Japan.

Tatsuya M. Ikeda1, W. John Rogers2, Gerard Branlard3, Roberto J.

Peña4, Silvia E. Lerner5, Adriana Arrigoni5, Wujun Ma6, Rudi Appels6,

Odean Lukow7, William Hurkman8, Marie Appelbee9, Mike Sissons10,

Jose M. Carrillo11 and Zhonghu He12

2CIISAS, CICPBA-BIOLAB AZUL, Facultad de Agronomía, Azul, UNCPBA,

Argentina. CONICET INBA -CEBB-MdP. 3INRA Station d'Amelioration des Plantes, Clermont- Ferrand, France.

4CIMMYT Mexico. 5CRESCAA, Facultad de Agronomía, Azul, UNCPBA, Argentina.

6Western Australia Department of Agriculture and Food, State Agriculture

Biotechnology Center, Murdoch University, Murdoch, Australia. 7Agriculture and Agri-Food Canada, Cereal Research Centre, Winnipeg,

Canada. 8USDA Agricultural Research Service, Western Regional Research Center,

Albany, USA. 9 South Australian Research and Development Institute, Adelaide and

LongReach Plant Breeders, Lonsdale, Australia. 10Tamworth Agricultural Institute, Calala, NSW, Australia.

11Unidad de Genética, ETSIA, Madrid, Spain. 12Institute of Crop Science, National Wheat Improvement

Center/The National Key Facility for Crop Genetic

Resources and Genetic Improvement, Chinese Academy of

Agricultural Sciences, Beijing, and CIMMYT, China Office,

Beijing, China.

But technical difficulties

in allelic identification due

to the complexity of the

protein profile produced

by each cultivar and the

use of different

nomenclature systems in

different laboratories has

historically interfered with

information exchange

between research groups

The current

contribution

summarises

progress made by

this group and seeks

to comment on

remaining challenges

Plus aims to place the findings in the

context of the Wheat Gene Catalogue...

In the current wheat gene

catalogue (McIntosh et

al., 2008 and annual

supplements), how many

alleles are there at each

of the three Glu-3 loci?

0,00 0,24 0,48 0,72 0,96

ACA 201B. Aguará

B. ChacareroB. Pingo

Las Rosas INTAACA 302

ACA 304B. Mejorpan

B. PonchoBio 2001

Bio 3001B. Norteño

ACA 303ACA 315

ACA 601ACA 301

B. RanquelCoop. Nahuel

PI CauquenB. Napostá

B. GuatimozinB. Malevo

Coop. LiquénCoop. Nanihue

B. ArrayánB. Farol

B. CharrúaB. Panadero

Baguette 21ACA 223

B. OmbúB. Puelche

Bio 1001K. Escorpión

K. ZorroPI Oasis

Bio 1002ACA 801

ZorzalCronox

OnixSirirí

PI EliteBio 3004

K. DragonK. Gavilán

PI Don UmbertoPI Federal

PI MileniumT. Nevado

PI GranarPI Real

K. CaciqueINIA Condor

INIA Pus 14K. Martillo

T. ChapelcoBio 1000

K. Don EnriqueINIA Tijetera

Agrovic 2000B. Baqueano

B. ChambergoK. Guerrero

PI GauchoBio 2000

K. CastorK. Tauro

PI ImperialACA 901

B. ManantialK. Capricornio

K. ChajáK. Flecha

B. ArrieroBio 2002

GreinaK. Estrella

B. HalcónB. Raudal

K. ProteoBio 1003

CentinelaB. Pronto

K. JabalíB. Sureño

K. DelfinB. 75 Aniversario

B. PatacónB. Yatasto

Bio 3000Bio 3002

Pampa INTAPI Redomón

B. GuapoK. Sagitario

PI HuenpanPI Molinero

B. MatacoPI Amanecer

PI AlazánB. Bigua

PI Cinco cerrosPI Colibrí

Bio 3003K. Escudo

Baguette 13K. Volcan

INIA ChurrinchePI Puntal

B. BrasilBaguette 20

K. ChamacoBaguette 10

Baguette 11B. Guaraní

LonaPI Isla Verde

Baguette 12

Enough to allow

Lerner et al. (2009

Journal of Cereal

Science 49: 337–345)

to use them, along

with variation in the

HMW-GSs, to find 93

allelic combinations in

119 Argentinean

cultivars...

“The main ambiguities from these different classification

systems

can be summarized as follows: 1) at the Glu-A3 locus, both Glu-A3a and Glu-A3c were

reported for the same cultivar, and similarly, Glu-A3a,

Glu-A3b, Glu-A3c, Glu-A3d were reported to be identical

to Glu-A3e; 2) at the Glu-B3 locus, results differed for Glu-B3b and

Glu-B3g, and for Glu-B3f and Glu-B3g in the same

cultivars; 3) at the Glu-D3 locus, there was ambiguity between

Glu-D3a and Glu-D3c, and between Glu-D3a and Glu-

D3b in the same cultivars. As a consequence of these

problems, reports of correlations between certain allelic

forms of LMW-GS and quality parameters in common

wheat have often been contradictory .

In Australian cultivars (Gupta and Shepherd 1988; Gupta

et al. 1989b, 1990a and b, 1991, 1994; Metakovsky,

1990), for Rmax (Maximum dough resistance), the Glu-A3

alleles ranked b>d>e>c, the Glu-B3 alleles ranked

i>b=a>e=f=g=h>c and the Glu-D3 alleles ranked:

e>b>a>c>d. The allele b of both Glu-A3 and Glu-D3

seemed to be associated with more extensible wheats.

Cornish et al. (1993) found that the Glu-3 allelic pattern bbb

(at Glu-A3, Glu-B3 and Glu-D3, respectively) gave the best

extensibility, especially when combined with the Glu-1 pattern

bba (at Glu-A1, Glu-B1 and Glu-D1, respectively). Glu-3 bbc

also had excellent extensibility. They also concluded that Glu-

A3e was detrimental to extensibility by virtue of being null and

that Glu-B3 c,d and g had medium to weak dough properties.

They suggested that the best combinations for Glu-3 are bbb,

bbc and cbc.

In durum wheat, allele Glu-B3s (formerly Glu-B3b) encoding

subunits 8+9+13+16 and allele Glu-A3k (formerly Glu-A3b)

encoding subunit 5 are associated with poor quality

Branlard et al. (2001) also compared allelic effects on

quality parameters, finding that, for dough strength, the

rankings were as follows: at Glu-A3: a=d=f≥e, at Glu-B3:

b’≥d=c=c’=b=g>i>f≥j and at Glu-D3: a≥b=d=c. For

extensibility at Glu-A3: d=a=f≥e, at Glu-B3:

i≥b’≥c=c’=g>b=f=d>j, while, at Glu-D3, no significant

differences were found. Luo et al. (2001) found that, in New Zealand cultivars: (i) the Glu-

A3 alleles ranked: d>c=e, coinciding with Gupta et al (1990a) for

Rmax; (ii) the Glu-B3 alleles ranked: b>g, which coincides both

with Gupta (1990a) and Cornish (1993); and (iii) the Glu-D3

alleles ranked: b>a.

It can be seen that not all the published allelic rankings

are consistent, implying considerable further work is

needed to be able to clarify the situation...

In this collection of

cultivars, Glu-A3a, Glu-A3b,

Glu-A3c and Glu-A3f could

be readily distinguished

Difficult to distinguish Glu-

A3e (null) and Glu-A3f . Both

tended to be identified as

null.

AndGlu-A3d and Glu-A3g could only be distinguished by the gliadin encoded by Gli-A1o linked to

Glu-A3d

Glu-B3d, Glu-B3h and Glu-B3i each

carried slow bands not always

easy to distinguish

Glu-B3b almost coincided with

Glu-B3a, but Glu-B3b band

was usually lighter and

thinner

Glu-B3f could not be readily distinguished from Glu-B3g

Alleles

classified as

Glu-B3b, Glu-

B3g and Glu-

B3i were often

identified as

Glu-B3ab, Glu-

B3ac and Glu-

B3ad by 2DE

Again the

gliadins

can help

Bands can be faintly stained and not

always easy to distinguish, although

technical improvements have

often allowed discrimination of, for

example, Glu-D3a, Glu-D3b and Glu-D3d .

Recourse to 2DE, MALDI-

TOF or PCR is often required

For example, Glu-A3d and Glu-A3g (used gliadins in

1D) could be distinguished from each other by 2DE

Glu-B3a and Glu-B3b, difficult in 1D, can be distinguished in

2DE

Glu-D3c and Glu-D3l, could

be distinguished in 2DE

Nonetheless, there are alleles were not readily distinguished by this

method

Cause difficulties in

1D SDS-PAGE, but

can be distinguished

here.

Cause difficulties in

1D SDS-PAGE, but

can be distinguished

here.

Cause difficulties in

1D SDS-PAGE, but

can be distinguished

here.

Control: 8-hour day 20°C, 16-hour

night 16°C

Treatment: Starting 16 days after

anthesis, 8-hour day 35°C, 16-hour

day 20°C for three days

Stress In isogenic

lines for

LMW-GSs

subjected to

heat stress,

the allele

Glu-B3h

showed a

reduction of

14% in the

relative

quantity of

protein

detected in

SDS-PAGE

Control

In relative

terms only a

handful of

alleles have

been

assessed for

quality...

... and

therefore only

a true

collaboration

between

many groups

will provide

sufficient

resources to

allow all

allelic

variants to be

evaluated...

Questio

n:

Total Glu-

A3 50

Total Glu-

B3 30

Total Glu-

D3 13 Grand Total Glu-3

93

This is in preparation for

the production of the full

catalogue for publication

in the Proceedings of the

International Wheat

Genetics Symposium,

Yokohama, 2013