Euphytica 3 1 (1982) 677490

USE OF THE SDS-SEDIMENTATION TEST AND

SDS-POLYACRYLAMIDEGEL ELECTROPHORESIS
FOR SCREENING BREEDER’S SAMPLES OF
WHEAT FOR BREAD-MAKING QUALITY
JOHANNESH.E.MOONEN,AUKESCHEEPSTRAandARIS GRAVELAND

Institute for Cereals Flour and Bread TNO, P.O. Box 15,670O AA Wageningen, the Netherlands

Received3 november 1981

INDEX WORDS

Triticum aestivum, wheat, glutenin, SDS-sedimentation test, SDS-polyacrylamidegel-electrophoresis, bak-

ing quality selection.

SUMMARY

Gelprotein or SDS-insoluble gel-forming glutenin was isolated from wheat flour by extraction with an
aqueous 1.5% SDS solution. Remarkable intervarietal differences were observed both in amount and subun-
it composition of these proteins.
The amount of gelprotein and the SDS-sedimentation volume both proved to be good parameters for
the bread-making quality of wheat cultivars. A high correlation was observed between amount of gelprotein
and SDS-sedimentation volume. The amount of gelprotein was therefore tentatively assumed to be the
essential basis of the SDS-sedimentation test.
The subunit composition of the gelprotein was studied by SDS-PAGE after reduction of SS bonds by
mercaptoethanol. It was found that the average bread-making quality of wheat cultivars and progeny of
the cross Atlas 66 x Atys which possessedsubunits 3 and 10, coded for by chromosome lD, was significantly
higher than that of wheat samples possessing subunits 2 and 11, their allelic counterparts.

INTRODUCTION

Nowadays it is generally accepted that wheat proteins are responsible for the visco-
elastic and bread-making properties of a dough. However there is still much dispute
as to which protein fraction is involved (WALL, 1979).
Recently, wheat proteins were fractionated in our laboratory by suspending wheat
flour in 1.5% sodiumdodecylsulphate (SDS) solution and ultracentrifugating the sus-
pension (GRAVELAND et al., 1979). A gel layer, mainly consisting of SDS-insoluble
glutenin could be isolated (GRAVELAND et al., to be published). It will be referred to
as gelprotein. Remarkable differences between wheat cultivars both in quantity and
viscosity of the gel layer were observed.
In 1978 AXFORD et al. introduced the SDS-sedimentation test, a relatively simple
method for estimating bread-making quality of wheat cultivars. This led us to study
the relationship between the amount of the SDS-insoluble gelprotein and the SDS-
sedimentation volume and their correlations with bread-making quality.
It has been shown by PAYNE et al. (1979, 1981) and BURNOLJF& BOURIQUET (1980)
that the composition of the high-molecular weight subunits of glutenin can be related
677
 J. H. E. MOONEN, A. SCHEEPSTRA AND A. GRAVELAND

with bread-making quality. These observations could not be confirmed by LAWRENCE

& SHEPHERD (1980) and ZEHATSCHEK (1980). To elucidate this contradiction we investi-
gated the composition of the high-molecular weight subunits of gelprotein from 60
wheat cultivars by SDS-polyacrylamidegel electrophoresis (SDS-PAGE) and its rela-
tionship with bread-making quality. The investigation moreover included the parents
and progeny of a cross between a poor and a good bread wheat.

MATERIALSANDMETHODS

Wheat samples. Samples of 60 wheat cultivars, grown in 1979, were kindly given by
Koninklijk Kweekbedrijf en Zaadhandel D. J. van der Have at Rilland (NL).
The cultivars Atlas 66 and Atys and Fg progeny from a cross between these were
obtained from the Foundation for Agricultural Plant Breeding in Wageningen; they
had been grown on trial fields in 1979.

Baking test. Standard baking tests and micro-baking tests were performed as described
by SMAK (1972) and MEPPELINK (198 1) respectively.

SDS-sedimentation test. The SDS-sedimentation test was carried out according to Ax-
FORD et al. (1978).

Isolation of gelprotein. Wheat flour was fractionated as described by GRAVELAND et

al. (1979). After ultracentrifugation of the wheat flour suspension in 1.5% SDS, the
gel layer was scraped off and washed with ethanol and water. The resulting white
precipitate (gelprotein) was lyophilized.

SDS-PAGE. SDS-PAGE was carried out with the buffer system described by laemmli
(1970). Gelprotein (10 mg/ml) or total flour (40 mg/ml) was suspended in a buffer
of 0.062 M Tris-HCl (pH 6.8) 2% SDS and 5% mercaptoethanol. After shaking for
2 h at room temperature the suspensions were boiled for 2 min. After centrifugation
(2000 g) for 5 min at room temperature 10 ul gelprotein solution or 50 ul flour extract
was applied to a vertical polyacrylamide slabgel (180 x 140 x 2.7 mm) which consisted
of a 4% (w/v) acrylamide, 0.107% (w/v) b isacrylamide stacking gel (20 mm) and a
10% (w/v) acrylamide, 0.267% (w/v) bisacrylamide separation gel (120 mm). The elec-
trophoresis was carried in a Pharmacia GE 2/4 system at room temperature, the elec-
trophoresis buffer was cooled with tapwater. Two slabgels were run with 30 mA for
30 min, and 60 mA overnight. Proteins were fixed and stained with 0.2% Coomassie
Brilliant Blue R 250 in HzO/MeOH/HAc (45/45/10; v/v/v) at 60°C for 1 h and des-
tained with HzO/MeOH/HAc (175/10/l 5; v/v/v) at 60 “C overnight.

Identification of the gelprotein subunits. After SDS-PAGE of gelprotein three groups

of subunits, classified as A, B and C could be distinguished. As pointed out in later
under ‘characterization’ only the A subunits were considered to be important for
bread-making quality of wheats.
The A subunits of the gelprotein were numbered by increasing mobility, the A su-
bunit with the lowest mobility having number 1. As a reference Chinese Spring pos-
sessesthe A subunits 2, 5, 8 and 11.
678 Euphytica 31 (1982)
 SCREENING FOR BREAD-MAKING QUALITY IN WHEAT

Statistics. For groups of cultivars or progeny with the same A subunits the average
loaf volume was calculated. Any differences in the average loaf volumes were tested
for significance with Student’s t test. At a probability level of 5% (P < 0.05) the differ-
ence was regarded as ‘probably significant’, at P < 0.01 the difference was ‘significant’
and at P < 0.001 ‘highly significant’.

RESULTSANDDISCUSSION

Correlation between amount of gelprotein, SDS-sedimentation volume and loaf volume.

We studied the relationships between protein percentage of flour, amount of gelprotein
in flour and SDS-sedimentation volume of whole-meal flour on the one hand, and
loaf volumes as determined by standard baking tests with flour on the other hand.
The results are shown in Fig. 1. It appears that both the amount of gelprotein and
SDS-sedimentation volume are good parameters for estimating bread-making quality.
The high correlation coefficient we calculated between SDS-sedimentation volume
and loaf volume (r = 0.87; P < 0.001) is in agreement with findings of AXFORD et
al. (1978, 1979). From the data of Fig. 1 we could expect that there also existed a
strong correlation between the amount of gelprotein and SDS-sedimentation volume.
This was confirmed: while the correlation coefficient between total protein and SDS-
sedimentation volume was r = 0.48 (P < 0.05) that between gelprotein and SDS-sedi-
mentation volume was much more pronounced: r = 0.95 (P < 0.001; see Fig. 2).
These correlation coefficients form a theoretical base for the SDS-sedimentation
test which was developed by AXFORD et al. (1978, 1979) and criticized by BOLLING
& M~NZING (1980). If wheats differ in their SDS-sedimentation volume they actually
differ in the amount of gelprotein or SDS-insoluble glutenin.
Characterization of gelprotein by SDS-PAGE. SDS-PAGE electropherograms of gel-
protein after reduction with mercaptoethanol from 10 European wheat cultivars are
shown in Fig. 3. In accordance with results of PAYNE et al. (1979) the subunits of
the gelprotein can be divided in three groups: group A with molecular weights over
70,000 d, group B with molecular weights between 40,000 and 50,000 d, and group
C with molecular weights between 30,000 and 40,000 d. The bands with varying intensi-
ty seen in the region with apparent molecular weights of about 68,000 d are an impurity
in the gelprotein preparation. They belong to the so-called glutelins according to GRA-
VELAND et al. (to be published).
Clear varietal differences in mobility of the subunits can be observed in all groups.
From the work of PAYNE et al. (1979, 1981) and BURNOUF & BOURIQUET(1980) we
know that differences in mobility of the A subunits may be related with differences
in baking quality between wheat cultivars.
We discovered that the A subunits were clearly visible already in total flour extracts;
this conveniently obviated the necessity of preparing gel-protein. This is shown in
Fig. 4.
Characterization of the A subunits from 60 wheat cultivars; relationship with bread-
making quality. The composition of the A subunits of 60 wheat cultivars was deter-
mined by SDS-PAGE of total flour extracts. The subunits were numbered by increas-
ing mobility, the subunit with the lowest mobility (i.e. the highest molecular weight)
Euphyiica 31 (1982) 679
 J. H. E. MOONEN, A. SCHEEPSTRA AND A. GRAVELAND

A I I I I I
v 20 25 30 35 40
V. PROTEIN ( ‘Iour 1 GELPROTEIN ( mg/ g flour )

700 -

:
0
;
600 -
Gl
8
<
E

w Fig. 1. Correlations between loaf volume

f 500 -
and protein content of the flour, amount
of gelprotein and SDS-sedimentation test.
‘“4 450 Open circles: wheat cultivars with A sub-
I! units 3 and 10. Closed circles: wheat cul-
I tivars with A subunits 2 and I 1.The wheat
I I I I I I
I Al
40 50 60 70 80 90 100 varieties used in this experiment are indi-
SDS-SEDIMENTATION VOLUME ( ml ) cated in Table 1 by *.

having no. 1. Within the 60 samples a total of 11 A subunits could be distinguished.

In Table 1 the wheat cultivars are arranged in groups with identical subunits. A wheat
cultivar appears to possess either subunits 3 and 10 (group I) of 2 and 11 (group II).
Statistic analysis shows that the difference in average loaf volume between group
I and group II is highly significant (Table 2).
If we compare our results with those of PAYNE et al. (1980, 1981) taking Chinese
Spring as a reference, we may assume that the subunits 3, 10 and 2, 11 are allelic
counterparts of each other, and are controlled by the long arm of chromosome 1D
(BIETZ et al., 1975).

680 Euphytica 31 (1982)

 SCREENING FOR BREAD-MAKING QUALITY IN WHEAT

- QOt

I I I I I
10 11 12 13 14
V. PROTEIN ( flour ) GELPROTEIN ( mg g flour )
I
Fig. 2. Correlation between SDS-sedimentation volume and protein content and amount of gelprotein.
For symbols see legends Fig. 1.

Recently PAYNE et al. (1979, 1981) have suggested that the presence of subunit 1
in wheat cultivars improves bread-making quality. However as shown in Table 2 the
difference in loaf volume between the cultivars with subunit 1 and those without sub-
unit 1 is not significant. Moreover, comparison of group I, 1 and group I, 2 shows that
the addition of subunit 1 did not improve bread-making quality.
However addition of subunit 8 (group I, 1 versus group I, 3) and replacement of
subunit 5 by subunits 4 and 8 (group I, 1 versus group I, 5) improved bread-making
quality significantly (Table 2). The difference between group I, 2 and I, 5 was not
significant; neither were the differences between the subgroups of group II (data not
shown).

Relationship between the A subunits in progeny of the cross Atlas 66 x Atys and bread-
making quality. An elegant method to establish whether the division between wheat
varieties made in the previous section is meaningful in respect to bread-making quality,
is to analyse the progeny of a cross between wheat varieties of which one parent has
the subunits 3 and 10 and the other parent the subunits 2 and 11.
In Fig. 5 SDS-PAGE electropherograms are shown of Fg progeny lines of the cross
Atlas 66 x Atys. Atys contains the subunits 1, 3, 5, 8, 10 and Atlas the subunits
2, 6, 7, 11. Fig. 6 indicates that no relationship exists between protein content of the
flour and loaf volume as determined by the micro-baking test. Besides it is shown
in Fig. 6 which lines possess the subunits 3 and 10, subunits 2 and 11, or the combina-
tion of these subunits. The difference in bread-making quality between group I and
group II proved to be significant (Table 3).
Euphytica 31 (1982) 681
 J. H. E. MOONEN, A. SCHEEPSTRA AND A. GRAVELAND

Fig. 3. SDS-PAGE of isolated gelprotein from 10 European wheat cultivars. Lane 1, Sicco; 2, Bastion;
3. Adonis; 4, Okapi; 5, Tundra; 6, Marksman; 7, Carimulti; 8, Kormoran; 9, Hardi; 10, Rivoli; 11, Calibra-
tion proteins, phosphorylase b (97,000), serum albumin (68,000), ovalbumin 43,000) and carbonic anhydrase
(30,000).

SDS-PAGE of single kernels of the F8 lines that possessed in the flour the combina-
tion of A subunits 2,3, 10 and 11 showed that these lines were still impure. The single
kernels possessed either the combination 2, 11 or the combination 3, 10 (results not
shown).
Again the presence of subunit 1 was not accompanied with enhanced bread-making
quality. Neither was there any significant difference in bread-making quality between
Fg lines possessing the subunits 6 and 7 and those possessing the subunits 5 and 8.
In the subgroups of group I and group II no further significant correlations between
subunits and bread-making quality could be found (Table 3).

GENERAL DISCUSSION

The usefulness of the SDS-sedimentation test as an estimate of bread-making quality

(AXFORD et al., 1979; BOLLING & M~~ZING, 1980) is disputed. We established in this
work that a strong correlation exists between the amount of gelprotein and the SDS-

682 Euphytica 31 (1982)

 SCREENING FOR BREAD-MAKING QUALITY IN WHEAT

Fig. 4. SDS-PAGE of isolated gelprotein and total flour extracts from 4 European wheat cultivars. Lane
1, Marksman flour; 2, Marksman gelprotein; 3, Tundra four; 4, Tundra gelprotein; 5, Bastion flour; 6,
Bastion gelprotein; 7, Sicco flour; 8, Sicco gelprotein.

sedimentation volume (r = 0.95; P < 0.001). Both properties give a rather good indica-
tion of the bread-making quality of wheat cultivars (Fig. 1 and 7) (Fig. 7 is a graphic
representation of Table 1).
However, we should remark that the SDS-sedimentation test was insufficiently ac-
curate in our hands to reveal existing differences in bread-making quality between
the subgroups of group I (data not shown).
It has recently been shown in our laboratory that the SDS-insoluble gelprotein be-
comes SDS-soluble by the action of a redox system present in wheat flour, when a
dough is mixed in the presence of oxygen (GRAVELAND et al., 1980). It is conceivable
that this redox system is also active during the extraction of flour proteins with SDS.
As indicated by GRAVELAND et al. (1979) both mechanical stress and elevated tempera-
tures could activate this redox system, thereby having a negative effect on the amount

Euphytica 31 (1982) 683

 J. H. E. MOONEN, A. SCHEEPSTRA AND A. GRAVELAND

Table I. A subunits, loaf volumes and SDS-sedimentation volumes of 60 wheat cultivars.

Group I A subunits 3, 10 Group II A subunits 2, 1 I

group 1, 1 (3, 5, IO) group II, I (2, 5, 11)

Kormoran* W D 660 96 Hardi* W F 646 96
Nimbus* W D 608 70 Cora* W B 609 68
Kolibri S D 562 79 H864-3 W B 594 69
Tina S B 519 52 Okapi* W D 587 70
Bayard S F 631 91 Arminda* W NL 539 69
Orca S NL 507 65
group I, 2 (1, 3, 5, 10)
Carimulti* W D 501 65
Echo S AUS 731 94
Sicco* S NL 691 96 group II, 2 (2, 5, 8, 11)
Hermes S D 619 76 Carisuper W D 576 80
Famos S D 669 70 Karamu S NZ 544 56
Janus* S E 665 88
group II, 3 (2, 5, 9, 11)
Max S 649 66 Rivoli* W F 591 71
Rongotea S NZ 635 65
Melchior S NL 540 65
Disponent* W D 633 64
Kaspar* S NL 513 54
Arawa S NZ 618 70
Toro S NL 549 75
group II, 4 (1, 2, 5, 11)
group I, 3 (3, 5, 8, 10)
Manella W NL 585 64
Newana
Blonda ; NL 521
580 45
66
Era E USA 750
726 97
83
Adonis*
Adam W A 679 95
Fortuna S USA 663 71 group II, 5 (I, 2, 5, 9, 11)
Pratos S F 662 87 Bastion* S NL 583 70
Hatri S DDR 629 65
group II, 6 (2, 6, 7, 11)
group I, 4 (1, 3, 5, 8, 10) Prinqual
Azur S F 650 17 Sappo* ; F 691
697 93
95
group I, 5 (3,4, 8, 10) group II, 7 (1,2, 6, 11)
Oroua S NZ 736 90 Olympic S AUS 576 65
Sonett s s 719 96
Duellant* W group II, 8 (2,4, 8, 11)
D 684 91
s s 688 84
Magnus
Hilgendorf ; S
NZ 658
587 87
67
Sarah W DK 579 63
Norda W NL 536 47
Marksman* W GB 531 72
Durin* W GB 499 64
Colombo W B 544 53
Tumult* W NL 495 49
Tundra* W E 487 44
Magister W 462 51
Holme w s 646 70
Hildur w s 641 72
groupII,9(1,2,4,8, 11)
Kopara S NZ 553 67
group II, 10 (2, 4, 7, 8, 11)
Highbury S GB 614 84
group II, 11 (2,4, 11)
Winnetou W DDR 508 40

The table indicates from left to right variety, type of wheat (summer or winter), country of origin, loaf
volume standard baking test (ml. 100 g flour -I), SDS sedimentation volume (ml. 6 g meal-t). The cultivars
indicated with * are used in the experiments of Fig. 1 and Fig. 2.

684 Euphytica 31 (1982)

 SCREENING FOR BREAD-MAKING QUALITY IN WHEAT

Fig. 5. SDS-PAGE of Fg progeny from the cross Atlas 66 x Atys.

Lanes 1 + 6 and 9 -+ 14 Fg progenies
7 Atys
8 Atlas 66 685
Table 2. Correlations between the presence of A subunits and loaf volume.

(Sub)Groups A subunits compared Probable Average loaf volumes Significance of higher

compared chromosomal loaf volumes for a
a B control G( /I subunits

I “S II 3, IO 2, II ID 656 f 57 (25) 572 + 61 (35) t58 = 5.44; P iO.001

varieties varieties
I vs 1 IA 622 k 58 (17) 601 k 77 (43) t58 = 1.02; P < 0.5
with without
vs
subunit I subunit I present absent
1, 2 vs I,1 1, 3, 5, 10 vs 3, 5, 10 IA 653 k 50 (10) 596 f 56 (5) t13 = 2.00; P < 0.1
1, 3 vs I,1 3, 5, 8, IO vs 3, 5, 0 IB 685 f 45 (6) 596 + 56 (6) t9 = 2.94; P < 0.02
1. 5 vs I,1 3,4,8, IO vs 3, 5, IO 1B 713 f 27 (3) 596 k 56 (5) t6 = 3.34; P < 0.02
1, 5 vs I, 2 3, 4, 8, IO vs I, 3, 5, IO 18, IA 713 + 27 (3) 653 + 50 (IO) tll = 1.94; P < 0.1

Table 3. Correlations between inheritance of A subunits and loaf volumes in Fg progeny of the cross Atlas 66 x Atys.

A subunits compared Probable Mean loaf volumes Significance of higher

chromosomal loaf volumes for a
cf. B control u B subunits

3, IO vs 2, II ID 501 f 40(17) 452 + 47 (I I) t26 = 2.94; P < 0.01

I present vs I absent IA 487 t 35 (25) 480 &- 67(ll) t34 = 0.57; P > 0.05
6, 7 VS 5, 8 IB 493 f 40 (13) 477 f 56(14) t25 = 0.85; P < 0.5
3, IO vs I, 3, IO IA 522 k 40 (6) 491 f 37(ll) tl5 = 1.61; P < 0.2
3, 5, 8, IO vs 3,6,7,10 IB 511 + 29 (7) 5llk39 (8)
1.2. 11 vs 2, II IA 471 k 31 (6) 429 f 56 (5) t9 = 1.58; P < 0.2
2, 6, 7, 11 vs 2,5,8, II IB 468k21 (4) 443 + 57 (7) t9 = 0.83; P < 0.5
 SCREENING FOR BREAD-MAKING QUALITY IN WHEAT

0 0
A0
00
0
atys
0 o" A

0 02. .
A. . l .
A
8.0°

A oA Oaths 66
0
.
0

I .

In I I I .1 I I
” 12 13 14 15 16 17

*lo PROTEIN IN FLOUR

Fig. 6. Correlation between loaf volume and protein content of the flour of F6 progeny from the cross
Atlas 66 x Atys.
Open circles : Fg progeny with A subunits 3 and 10
Closed circles : Fg progeny with A subunits 2 and 11
Triangles : Fg progeny with A subunits 2,3, 10 and 11

of SDS-insoluble glutenin. Reproducible mechanical shaking in a temperature-con-

trolled bath or, alternatively a total blocking of the redox system could therefore im-
prove the usefulness of the SDS-sedimentation test.
The involvement of the A subunits in the bread-making quality of wheats is well
documented (ARAKAWA et al., 1977; PAYNE & CORFIELD, 1979; WALL, 1979). Several
research groups (ORTH & BUSHUK, 1974; BIETZ et al., 1975; BURNOUF & BOURIQUET,
1980; LAWRENCE & SHEPHERD, 1980; PAYNE et al., 1980) have established that the
A subunits of wheat glutenin are controlled by the homoeologous group 1 chromo-
somes. Comparing their evidence with our results we assume that subunit 1 is con-
trolled by chromosome lA, subunits 2, 3, 10 and 11 by chromosome lD, and the
other subunits by chromosome 1B. In accordance with the results of PAYNE et al.
(1981) we found a significantly better bread-making quality both in wheat cultivars
and in Fg progeny of the cross Atlas 66 x Atys, containing the 1D controlled subunits
3 and 10 compared with samples containing the 1D controlled subunits 2 and 11 (Table
2 and 3). Furthermore the addition of subunit 8 to the subunits 3, 5 and 10, or the
replacement of subunit 5 (in combination with the subunits 3 and 10) by subunits
4 and 8 improves bread-making quality (Table 2). In contrast to PAYNE et al. (1979)
we were not able to confirm the positive involvement of subunit 1 in bread-making
quality.
One might conclude from Figs 1 and 2 that the higher bread-making quality of
cultivars of group I is caused by a higher protein content in the flours. However, the
average protein content of the flours of group I was 12.84 If: 0.92% and that of group
II 12.28 f 1.32x, the difference being not significant (P < 0.1). Besides it is shown
Euphytica 31 (1982) 687
 J. H. E. MOONEN, A. SCHEEPSTRA AND A. GRAVELANO

800
1
;2,700-
8
<
-i

lr!
2
?a00 -
%
2

500-

400 -
I I 1 1 I 1 I I
40 50 60 70 60 90 100

SDS- SEDIMENTATION VOLUME ( ml/69 meal 1

Fig. 7. Correlation between loaf volume and SDS-sedimentation volume of 60 wheat cultivars.
Open circles : cultivars with A subunits 3 and 10
Closed circles : cultivars with A subunits 2 and 11
The dotted circles indicate that the average SDS-sedimentation volume of cultivars with A subunits 3 and
10 is significant higher than the average SDS-sedimentation volume of cultivars with A subunits 2 and
11.

in Fig. 6 that the higher bread-making quality of the Fg progeny from Atlas 66 x
Atys possessing subunits 3 and 10, was definitely not caused by higher protein contents
of their flours.
The higher bread-making quality of cultivars with subunits 3 and 10 has to be ex-
plained by the fact that the cultivars of group I contained more SDS-insoluble glutenin
and higher SDS-sedimentation volumes than did the cultivars of group II. The mean
SDS-sedimentation volume of group I was 80.2 f 13.0 ml and that of group II 66.9
f 14.2 ml (see also Fig. 7). From Fig. 2 we can deduce that these values correspond
with 32.1 f 3.5 and 28.5 + 3.8 mg gelprotein/g flour respectively. The difference
in average amount of gelprotein between group I and group II was highly significant
(P < 0.001). To explain this we could assume that the A subunits 3 and 10 form more
stable and/or more numerous disultide bonds with the other glutenin subunits than
do the A subunits 2 and 11.
688 Euphyiica 31 (1982)
 SCREENING FOR BREAD-MAKING QUALITY IN WHEAT

The results of this study demonstrate that determination by SDS-PAGE of the A

subunit composition of wheat gelproteins, both in released cultivars and new lines
of a breeding programme, is a relatively simple test for assessing bread-making quality.

ACKNOWLEDGEMENT

This investigation was carried out with financial support of the Stichting Nederlands
Graan-Centrum, Wageningen.

NOTE

In recent work, the presence of a further subunit 2* was established in some wheat
cultivars, in accordance with the studies of G. J. LAWRENCE & K. W. SHEPHERD (Austr.
J. Biol. Sci 33 (1980) 221-233) and P. I. PAYNE, L. M. HOLT & C. N. LAW (Theor.
Appl. Genet. 60 (1981) 229-236). This subunit bears a positive effect on bread-making
quality. In a further paper, the identification of this subunit and its use in screening
breeder’s samples for baking quality will be described.

REFERENCES

ARAKAWA, T., M. YOSHIDA,H. MORSHITA,J. HONDA&D. YOHEZAWA, 1977. Relation between aggregation
behavior of glutenin and its polypeptide composition. Agric. Biol. Chem. 41: 995-1001.
AXFORD, D. W. E., E. E. MCDERMOTT& D. G. REDMAN, 1978. Small-scale test of bread-making quality.
Milling Feed and Fertilizer 66: 18-20.
AXFORD, D. W. E., E. E. MCDERMOTT & D. G. REDMAN, 1979. Note on the sodium dodecyl sulphate
test of bread-making quality: Comparison with Pelshenke and Zeleny tests. Cereal Chem. 56: 582-584.
BIETZ,J. A., K. W. SHEPHERD& J. S. WALL, 1975. Single-kernel analyses of glutenin: use in wheat genetics
and breeding. Cereal Chem. 52: 513-532.
BOLLING, H. & K. M~~NZING,1980. Die Verwendung der SDS-Sedimentation zur Beurteilung von Backwei-
zen. Getreide, Mehl und Brot 34: 611.
BURNOUF,T. & R. BOURIQUET,1980. Glutenin subunits of genetically related European hexaploid wheat
cultivars: their relation to bread-making quality. Theor. Appl. Genet. 58: 107-I 11.
GRAVELAND, A., P. BONGERS& P. BOSVELD,1979. Extraction and fractionation of wheat flour proteins.
J. Sci. Food Agric. 30: 71-84.
GRAVELAND, A., P. BOSVELD, W. J. LICHTENDONK &J. H. E. MOONEN, 1980. Superoxide involvement in
the reduction of disullide bonds of wheat gelprofeins. Biochem. Biophys. Res. Commun. 93: 1189-l 195.
GRAVELAND, A., P. BOSVELD,W. J. LICHTENDONK,J. H. E. MCXNEN & A. SCHEEPSTRA,Extraction and
fractionation of wheat flour proteins. II. J. Sci. Food Agric. to be published.
LAEMMLI, U. K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage
T4. Nature 227: 68&685.
LAWRENCE,G. J. & K. W. SHEPERD,1980. Variation in glutenin subunits of wheat. Aust. J. Biol. Sci. 33:
221-233.
MEPPELINK, E. K., 1981. EinsatzmGglichkeit des Microbackversuches in der Weizenziichtung. Getreide,
Mehl und Brot. 35: 107-109.
ORTH, R. A. & W. BUSHUK, 1974. Studies on glutenin VI. Chromosomal location of genes coding for
subunits of glutenin of common wheat. Cereal Chem. 51: 118-126.
PAYNE, P. I. & K. G. CORFIELD, 1979. Subunit composition of wheat glutenin proteins, isolated by gel
filtration in a dissociating medium. Planta 145: 83-88.
PAYNE, P. I., K. G. CORFIELD &J. A. BLACKMAN, 1979. Identification of a high-molecular-weight subunit
of glutenin whose presence correlates with bread-making quality in wheats of related pedigree. Theor.
Appl. Genet. 55: 153-159.

Euphyrica 31 (1982) 689

 1. H. E. MOONEN, A. SCHEEPSTRA AND A. GRAVELAND

PAYNE, P. I., L. N. LAW & E. E. MUDD, 1980. Control by homoeologous group I chromosomes of the
high-molecular-weight subunits of glutenin, a major protein of wheat endosperm. Theor. Appl. Genet.
58: 113-120.
PAYNE, P. I., K. G. CORFIELD, L. M. HOLT & J. A. BLACKMAN, 1981. Correlations between the inheritance
of certain high-molecular-weight subunits of glutenin and bread-making quality in progenies of six crosses
of bread wheat. J. Sci. Food. Agric. 32: 51-60.
SMAK, C. 1972. New approach to determine the browness of bread crust. Correlation between crust colour
and protein content. Cer. Chem. 49: 554-560.
WALL, J. S., 1979. The role of wheat proteins in determining baking quality. In: D. L. LAIDMAN & R.
G. WYN JONES (Eds), Recent advances in the biochemistry of cereals. Academic Press, pp 275-311.
ZEHATSCHEK, W., 1980. Das Elektrophoresemuster von Weizenglutenin bei Sorten mit unterschiedlichem
Qualitltsniveau. Getreide, Mehl und Brot 34: 239-243.

690 Euphytica 31 (1982)