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

1. LOW MOLECULAR WEIGHT M.T. LABUSCHAGNE
and A. VAN BILJON
GLUTENIN SUBUNITS
AND GLIADINS IN 137 Department of Plant
Sciences, University of
DOUBLE HAPLOID WHEAT the Free State, P.O. Box
LINES DERIVED FROM A 339, Bloemfontein
CROSS BETWEEN 9300, South Africa
Email:
KARIEGA AND AVOCET labuscm@ufs.ac.za
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za

2. INTRODUCTION
theWheat gliadins are a highly polymorphic essential amino
protein of maize is deficient in two group of seed
acids, lysine and tryptophan proteins
storage
They are controlled by Gli loci on the short arms of
chromosomes 1 and 6
γ and ω = 1A, 1B, 1D Gli-A1, Gli-B1, Gli-D1
α and β = group 6 chromosomes, Gli-A2, Gli-B2, Gli-D2

3. From Shewry and Lookhart (2003)

4. Genes encoding LMW GS (Glu-3) are linked to genes (Gli-1)
encoding ω and γ gliadins
Gliadin extract gives ω5, ω1,2, α and γ gliadins
Glutenin extract gives glutenin bound ω extracts and HMW
and LMW subunits

5. Most wheat cultivars show similar overall patterns of
gliadin and glutenin subunits with characteristic regions for
each type

6. AIM OF THE
AIMSTUDY
OF STUDY
 To study low
To study low
molecular weight
molecular weight
glutenins and
glutenins and
gliadins in a DH
gliadins in a DH
population of a
A
population of a
cross between
cross between
Kariega and Avocet
Kariega and Avocet

7. Materials and methods
Double haploid wheat population, developed with wheat x maize
crosses using wheat x maize crosses
Avocet x Kariega: 137 DH’s developed from F2 material
Kariega = hard red South African bread wheat with excellent quality
Avocet = standard white Australian spring wheat
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za

8. Materials and methods
Protein extraction for RP-HPLC using method of Wieser et al. (1998)
First gliadin extraction with 70% ethanol
Glutenin extraction with 50% propan-1-ol, urea and DTT and 4-
vinylpyridine
RP-HPLC with C18 column
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za

9. Results
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za

10. ωωω ωωα ωωγ
Gliadin profile of Avocet
ωωω ωωα ωωγ
Gliadin profile of Kariega

11. GLIADIN PEAK AREA (%) OF PARENTS AND THE DH
POPULATION
Name P20.8 P21.4 P22.3 P23 p24 p25.1 p26 p265 p27 p275 p28.6 p31 p345 p36
Avocet 2.91 6.13 0.00 0.00 2.4 0.00 13.12 10.35 4.25 8.00 2.66 18.47 6.89 1.27
Kariega 3.99 3.63 1.94 1.3 2.3 2.65 8.67 7.89 4.02 7.49 0.00 18.49 5.03 1.33
DH 3.44 3.86 1.28 1.77 2.36 0.900 13.16 11.29 5.13 6.69 2.59 16.57 6.77 1.74
ω α γ

12. Gliadins
 ω peak 22.3 was absent in Avocet but was
present in 50 of 137 DH’s
 ω peak 23 was absent in Avocet but was
present in 127 of 137 DH’s
 α peak 25.1 was absent in Avocet, but was
present in only 41 of 137 DH’s
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za
8/14/2012

13. Gliadins
 α peak 28.6 was absent in Kariega, but was
present in 118 of 137 DH’s
 α peak 26 was much higher in Avocet (13.12)
than in Kariega (8.67) but the DH average was
13.16, like the Avocet parent
 α peak 26.5 was higher for the DH average
(11.29) than either parent Avocet (10.35) and
Kariega (7.89)
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za
8/14/2012

14. Gliadins
 α peak 27.5 was lower for the DH average
(6.69) than for either parent Avocet (8) or
Kariega (7.49)
 The other peaks for DH averages were close
to the average of the two parents
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za
8/14/2012

15. Highly significant (P≤0.0001) correlations between
gliadins
Characteristic1 Characteristic2 Correlation
P20.8 (ω) P21.4(ω) -0.693
P20.8(ω) P22.3(ω) -0.432
P21.4 (ω) P22.3 (ω) 0.872
P22.3 (ω) P24 (α) 0.376
P22.3 (ω) P31 (α) -0.376
P23 (α) P36 (γ) 0.346
P25.1 (α) P26 (α) -0.886
P27.5 (α) P28.6 (α) -0.685

16. 0, 7+8, 2+12
Avocet glutenins
HMW glutenins
H HMW LMW glutenins
HM
2*, 17+18, 2+12
Kariega glutenins
HMW glutenins
HMW LMW glutenins
HMW
8/14/2012

17. LMW GLUTENIN PEAKS (% AREA) OF PARENTS AND THE DH
POPULATION
gl26 gl27 gl28.5 gl28.8 gl29.4 gl30 gl32 gl33.3
Avocet 6.72 37.05 5.17 3.63 3.60 6.06 3.22 4.03
Kariega 5.96 33.40 4.90 3.46 3.71 5.93 3.18 2.34
DH 9.79 28.97 4.24 4.03 3.34 5.34 3.98 5.41

18. LMW glutenins
 These subunits were more difficult to score
than the gliadins
 Peak 26 average of the DH was much higher
(9.79) than that of either Avocet (6.72) or
Kariega (5.96)
 Peak 27 average of the DH was much lower
(28.97) than either Avocet (37.05) or Kariega
(33.4)
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za
8/14/2012

19. LMW glutenins
 Peak 33.3 DH average (5.41) was much higher
than that of either Avocet (4.03) or Kariega
(2.34)
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za
8/14/2012

20. Highly significant (P≤0.01) correlations between LMW glutenins
Characteristic1 Characteristic2 Correlation
Peak 26 Peak 27 -0.372
Peak 28.8 Peak 29.4 -0.318
Peak 28.8 Peak28.5 0.243
Peak 28.5 Peak 29.4 -0.248
Peak 29.4 Peak 28.8 -0.318
Peak 28.8 Peak 30 -0.274
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za

21. Highly significant (P≤0.01) correlations between LMW glutenins and gliadins
Characteristic1 Characteristic2 Correlation
Gliadin peak 26.5 Glutenin peak 27 -0.229
Gliadin peak 27.5 Glutenin peak 28.5 0.253
Gliadin peak 25.1 Glutenin peak 28.5 -0.273
Gliadin peak 34.5 Glutenin peak 33 0.419
T: 051 401 9111 info@ufs.ac.zawww.ufs.ac.za

22. DISCUSSION AND CONCLUSIONS
8/14/2012

23. For the gliadins three peaks were absent in Avocet which
were in Kariega, and one was absent in Kariega which was
present in Avocet
Peak 22.3 absent in Avocet was present in 36% of the DH
population
Peak 23 absent in Avocet was present in 93% of DH population

24. Peak 25.1 absent in Avocet was present in 30% of DH
population
Peak 28.6 absent in Kariega was present in 86% of the DH
population

25. Peak 21.4 and 22.3 almost always occurred together and
were correlated r=0.872
Peaks 25.1 and 26 were highly negatively correlated (-0.886)
indicating that the presence of one lead to the supression of
the other
Peaks 20.8 and 21.4; and 27.5 and 28.6 were also highly
negatively correlated (-0.693 and -0.685) indicating that the
presence of one usually supressed the expression of the other

26. Eight clear LMW glutenin peaks were seen in all material
In four of the peaks the values were higher for the DH
population than that of either parent, and for the four other
peaks they were lower than for either parent
There were some significant correlations within the LMW
glutenins and between LMW glutenins and the gliadins, but
the values were very low

27. Both the glutenin and the gliadin peaks of the DH population
were not intermediate between that of the parents, indicating
some interaction between genes coding for these loci
The DH population expressed the peak missing in either parent
to a much higher extent than expected in two cases and much
lower than expected in two cases. One can speculate on the
reason for this

28. Thank you!