International Gluten Workshop, 11th; Beijing (China); 12-15 Aug 2012

1. 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

2. 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
1NationalAgriculture and Food Research Organization, Hiroshima, Japan.
2CIISAS, CICPBA-BIOLAB AZUL, Facultad de Agronomí Azul, UNCPBA,
a,
Argentina. CONICET INBA -CEBB-MdP.
3INRA Station d'Amelioration des Plantes, Clermont- Ferrand, France.
4CIMMYT Mexico.
5CRESCAA, Facultad de Agronomí Azul, UNCPBA, Argentina.
a,
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.

3. 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

4. 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...

5. In the current wheat gene
catalogue (McIntosh et Baguette 12 PI Isla Verde
Lona
Baguette 11 B. Guaraní
Baguette 10
K. Chamaco
al., 2008 and annual B. Brasil
INIA Churrinche
Baguette 13
Bio 3003
Baguette 20
PI Puntal
K. Volcan
K. Escudo
PI Colibrí
PI Cinco cerros
supplements), how many PI Alazán
B. Mataco
PI Huenpan
B. Bigua
PI Amanecer
PI Molinero
B. Guapo K.PI Redomón
Sagitario
alleles are there at each Pampa INTA
Bio 3000 Bio 3002
B. Yatasto
B. Patacón B. 75 Aniversario
K. Delfin
K. Jabalí B. Sureño
of the three Glu-3 loci? Centinela
K. Proteo
B. Halcón
Greina
B. Pronto
Bio 1003
B. Raudal
K. Estrella
Enough to allow B. Arriero
K. Chajá
Bio 2002
K. Flecha
B. Manantial K. Capricornio
ACA 901
PI Imperial K. Tauro
Lerner et al. (2009 K. Castor
PI Gaucho
B. Chambergo
Agrovic 2000
Bio 2000
K. Guerrero
B. Baqueano
K. Don Enrique INIA Tijetera
Journal of Cereal T. Chapelco
INIA Pus 14
K. Cacique
PI Granar
Bio 1000
K. Martillo
INIA Condor
PI Real
T. Nevado
Science 49: 337–345) PI Milenium
PI Don Umberto
K. Dragon
PI Elite
PI Federal
K. Gavilán
Bio 3004
Sirirí
Onix Cronox
to use them, along Zorzal
Bio 1002 ACA 801
PI Oasis
K. Zorro K. Escorpión
Bio 1001
B. Ombú B. Puelche
with variation in the Baguette 21
B. Charrúa
ACA 223
B. Panadero
B. Arrayán Coop.B. Farol
Nanihue
Coop. Liquén B. Malevo
HMW-GSs, to find 93 B. Guatimozin
PI Cauquen Coop.Napostá
B. Ranquel
ACA 601
B.
Nahuel
ACA 301
ACA 315
ACA 303 B. Norteño
allelic combinations in Bio 3001
B. Poncho
ACA 304
Las Rosas INTA
Bio 2001
B. Mejorpan
ACA 302
B. Pingo
B. Chacarero
119 Argentinean ACA 201 B. Aguará
0,00 0,24 0,48 0,72 0,96
cultivars...

6. “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 .

7. 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.

8. 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.
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
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...

9. 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.

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

11. 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

12. 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

13. Again the
gliadins
can help

14. 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

15. For example, Glu-A3d and Glu-A3g (used gliadins in
1D) could be distinguished from each other by 2DE

16. Glu-B3a and Glu-B3b, difficult in 1D, can be distinguished in
2DE

17. Glu-D3c and Glu-D3l, could
be distinguished in 2DE
Nonetheless, there are alleles were
not readily distinguished by this
method

18. Cause
difficulties in
1D SDS-
PAGE, but
can be
distinguished
here.

19. Cause
difficulties in
1D SDS-
PAGE, but
can be
distinguished
here.

20. Cause
difficulties in
1D SDS-
PAGE, but
can be
distinguished
here.

27. 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

28. In isogenic Stress Control
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

29. Questio
n:
... and
therefore only
a true
collaboration In relative
between terms only a
many groups handful of
will provide alleles have
sufficient been
resources to assessed for
allow all quality...
allelic
variants to be
evaluated...

33. Total Glu-
A3 50

36. Total Glu-
B3 30

37. Total Glu-
D3 13
Grand Total Glu-3
93

46. This is in preparation for
the production of the full
catalogue for publication
in the Proceedings of the
International Wheat
Genetics Symposium,
Yokohama, 2013