全 文 ：Vol. 30 , No. 6
pp. 544～547 　June , 2004
作 　物 　学 　报
ACTA AGRONOMICA SINICA
第 30 卷 第 6 期
2004 年 6 月 　544～547 页
Identification and Characterization of a Novel Wheat Line Containing
High Molecular Glutenin Subunits 5 + 12
WANG Tao1 , LI Zhu2Lin1 , REN Zheng2Long2 Ξ
(1 Chengdu Institute of Biology , Chinese Academy of Sciences , Chengdu 610041 , Sichuan ; 2 Sichuan Agricultural University , Plant Genetics and Breeding Key Labo2
ratory of Sichuan Province , Ya’an 625014 , Sichuan , China)
Abstract 　A new bread wheat line possessing high2molecular2weight glutenin subunits( HMW2GS) 5 + 12 was developed by
interspecific cross ( T. durum Desf . ×T. aestivum L. ) . The SDS2PAGE analysis demonstrated that this new germplasm
possessing HMW2GS 5 + 12 at Glu2D1 locus. Lines possessing HMW2GS 5 + 12 subunits were superior in many quality pa2
rameters , sedimentation value , wet gluten content , development time and stability time of farinograph as compared to their
siblines containing HMW2GS 5 + 10. This novel line had high yielding characters with multi2florets , and would be a poten2
tial genetic resource for bread wheat quality breeding.
Key words 　HMW2GS;Bread wheat quality ;Novel wheat line
具有高分子量谷蛋白亚基 5 + 12 的稀有小麦品系的鉴定和特性研究
王　涛1 　李竹林1 　任正隆2
(1 中国科学院成都生物研究所 ,四川成都 610041 ;2 四川农业大学植物遗传育种省级重点实验室 ,四川雅安 625014)
摘 　要 　采用种间杂交 (硬粒小麦 ×普通小麦)创造了具有高分子量谷蛋白 5 + 12 亚基的稀有小麦新品系 ,SDS2PAGE分
析表明 ,该品系的 Glu2D1 位点编码 5 + 12 亚基。其沉降值、湿面筋含量、粉质仪图形成时间、稳定时间等品质指标显著优
于具有优质亚基 5 + 10 的姊妹系 ,该小麦品系遗传上纯合 ,具有多花多粒的优良农艺性状。5 + 12 亚基稀有小麦新品系
的创造 ,将为小麦品质育种提供新的优质遗传资源。
关键词 　高分子量谷蛋白亚基 ;小麦品质 ;稀有小麦品系
中图分类号 :S512
　　The glutenin group of seed storage proteins in both
bread wheat ( Triticum aestivum L. ) and durum wheat
( Triticum durum Desf . ) , particularly the high molecular
weight glutenin subunits (HMW2GS) , are major determi2
nants in the bread baking and pasta making quality[1 ] . In
addition to HMW2GS compositions , low molecular weight
glutenin subunit 2 (LMW22) is also related with durum
wheat quality[6 ] . The HMW2GS are encoded by genes at
the Glu2A1 , Glu2B1 and Glu2D1 loci on the long arm of
chromosomes 1A , 1B and 1D , respectively[2 ] . Among
them , the 5 + 10 subunit encoded by the Glu2D1 locus
has a predominant effect in determining the baking quality
of wheat flours[3 ] . Hexaploid varieties containing novel
glutenin subunits , such as 5 + 12 and 1. 5 + 10 encoded
by Glu2D1 have been developed from crossing Triticum
turgidum ( 2 n = 28 , AABB ) and Triticum tauschii
(2 n = 14 , DD) , and the bread baking quality of flours
from synthetic wheat lines containing 5 + 12 glutenin sub2
units have been shown to be much better than other com2
bination lines with glutenin subunits , such as 5 + 10 or 2
+ 12 encoded by Glu2D1 [4 ] . The 5 + 12 subunit combi2
nation is rarely presented in cultivars of common
wheat [5～7 ]. We developed a new wheat line possessing
high2molecular2weight glutenin subunits 5 + 12 by inter2
specific cross ( T. durum Desf . × T. aestivum L. ) .
Here , we report the identification and preliminary qualityΞFoundation items :This research was supported by the grants from the Ministry of Science and Technology (2002AA207004) ; the Chinese Academy of Sci2
ences KSCX22SW2304 ,KSCX22120122204.
Biography :Wang Tao (1970 - ) , Male ,Associate Professor ,wheat breeder. Research projects including genetics and breeding of wheat quality , rust resis2
tance , and heterosis. E2mail : wangtao @cib. ac. cn
Received(收稿日期) : 2003206226 ,Accepted(接受日期) : 2003212221.

characterization of this novel wheat line containing the 5
+ 12 glutenin subunit .
1 　Materials and Methods
1. 1 　Plant Materials
The W7268272958 and W7268242301 , and sibline
W7268212168 line containing the 5 + 12 , and 5 + 10
glutenin subunits combination , respectively were
developed from durum wheat 779 (20) and bread wheat
81224 (N , 7 + 9 , 2 + 12) .
1. 2 　Methods
1. 2. 1 　SDS2PAGE 　　Standard SDS2PAGE technology
was used for HMW glutenin analysis[2 ] . The identification
of various HMW glutenin subunits was according to
Payne[8 ] using Chinese Spring as the control . The relative
quantity of HMW2GS were determined by Bio2Rad Model
GS2700 imaging desitometer.
1. 2. 2 　Determination of baking quality 　　Protein con2
tent , wet gluten content and Zeleny sedimentation value
were measured according to the standards of GB2905282 ,
GB/ T14608293 and AACC 56261A. Farinogragh and SDS
sedimentation value measurements were followed by
AACC54221 and AACC56263 standards.
1. 2. 3 　Determination of chromosome numbers and mei2
otic behaviors 　　Standard root tip method was followed
to determine the chromosome number. The meiotic behav2
iors were observed with pollen mother cells.
2 　Results
2. 1 　Determination of 5 + 12 subunits
Using Chinese Spring (N , 7 + 8 , 2 + 12) , 81224
(N , 7 + 9 , 2 + 12) , Chuanyu 12 (1 , 7 + 8 , 5 + 10) ,
the HMW glutenin subunits type in the W7268272958 line
was determined to have subunits 1 , 7 , 9 , 5 , and 12.
The subunits 1 , 7 + 9 and 5 + 12 are encoded by Glu2
A1 , 2B1 and 2D1 , respectively. Therefore , the W72682
72958 carries the subunits 5 + 12 at Glu2D1 (Fig. 1) .
2. 2 　Determination of cyto2genetic stability of 5 + 12
subunits in W7268272958
After the agronomic characters of the inter2specific
hybrid line W7268272958 were stabilized , we further de2
termined its cyto2genetic stability by determining the chro2
mosome number and meiotic behavior in W7268272958
Fig. 1 Lane 4 : W7268272958( 1 ,7 + 9 ,5 + 12) ;
Lane 5 : Chuanyu 12( 1 , 7 + 8 , 5 + 10) ;
Lane 6 : Chinese Spring ( N, 7 + 8 , 2 + 12)
and its sibline W7268212168 , parental durum wheat 799
(AABB ,2 n = 28) and 81224 (AABBDD ,2 n = 42) . The
chromosome numbers in W7268272958 and its sibline
W7268212168 lines were normal hexaploid (2 n = 42) and
stable. The meiosis in pollen mother cells was normal
too. Therefore , the W7268272958 line is homozygous ,
genetically and agronomically stable , and can be an ex2
cellent breeding stock which has very good agronomical
characteristics of 26 florelets , 144 kernels per spike , and
45. 6 g per 1 0002grain weight in the environment of
Chengdu.
2. 3 　Co2dominant inheritance of 5 + 12 subunits
The composition of HMW glutenin subunits in lines
81224 , W7268212168 , W7268272958 , and in F1 and F2
plants of crosses between W7268272958 and 81224 or
W7268212168 was analyzed by SDS2PAGE. There were
7 + 9 , 2 + 12 , and 1 , 7 + 9 ,5 + 12 subunits , in the 812
24 and W7268272958 lines respectively and the subunits
of 1 , 7 , 9 , 2 , 5 , 12 in the F1 plants of the cross between
the two lines. The co2existence of 2 and 5 subunits in the
hybrid indicated that the two subunits had co2dominant
inheritance since the genes coding 5 and 2 subunits were
located on the chromosome 1D. Similarly , in F1 plants of
the cross between W7268212168 containing 1 ,7 + 9 ,5 +
10 subunits and W7268272958 with 1 , 7 , 9 , 5 , 10 , 12
subunits , subunits 10 and 12 were also co2dominantly in2
herited. Therefore , subunits 5 and 12 , just as other sub2
units , are co2dominant .
SDS2PAGE analyses showed that F2 plants from
crosses between lines containing 5 + 12 and 2 + 12 had a
similar portion of the two parental types and the hybrid
type 2 + 5 + 12 (31∶30∶29) . However , F2 plants from
crosses between lines containing 5 + 10 and 5 + 12 pre2
545　6 期 WANG Tao et al . : Identification and Characterization of a Novel Wheat Line Containing High Molecular Glutenin Subunits 5 + 12 　　

sented an unusual segregation ratio , 49 (5 + 12 combina2
tion)∶17 (5 + 10) ∶7 (5 + 10 + 12) , which indicated a
different inheritance for 5 + 12 combination.
2. 4 　 Relative content of individual subunits in
various lines
Analyses of relative contents of individual subunits of
HMW2GS indicated that they were varied in different
wheat lines ( Table 1 ) . In Chinese Spring , 81224 ,
W7268212168 , W7268272958 , and W7268242301 lines ,
the relative contents of individual subunits were ranked
as : subunit 12 > 7 > 8 > 2 , 12 > 7 > 2 > 9 , 12 > 7 > 2 >
9 , 12 > 7 > 5 > 9 > 1 , and 12 > 7 > 5 > 9 > 1 , respec2
tively.
W7268272958 and W7268242301 lines were very
similar in the composition and relative contents of HMW2
GS subunits , but different significantly from W7268212
168 in the relative content of subunit 9. Additionally ,
W7268272958 (29. 66 %) and W7268242301 (30. 74 %)
compared to W7268212168 (23. 62 %) were 6. 04 % and
7. 12 % higher in the relative content of the subunit 7 re2
spectively , and W7268272958 (subunit 12 , 34. 96 %) and
W7268242301 ( subunit 12 , 33. 77 %) compared to
W7268212168 ( subunit 10 , 25. 49 %) 9. 47 % and
8. 28 % higher respectively in the relative content of the
y2type subunit at Glu2D1 locus.
2. 5 　The impact of the 5 + 12 subunits on the baking
quality of wheat flours
The impact of the 5 + 12 subunits on the baking
quality of wheat flours was assessed by comparing baking
quality parameters of flours from W7268272958 , it’s sib2
line W7268212168 and W7268242301 , and parental 81224
and durum wheat 779 lines (Table 2) . The W7268272958
and W7268242301 lines contained the 5 + 12 subunits ,
and had the higher SDS sedimentation values than their
siblines and parental lines.
The W7268272958 and W7268212168 lines had the
same HMW2glutenin subunits encoded at Glu2A1 and
Glu2B1 loci , but differed at the Glu2D1 locus. The two
lines have very similar agronomical properties because
they were F4 siblines. Thus , the flour quality difference
between these two lines was most likely due to the differ2
ence between the 5 + 12 and 5 + 10 subunits encoded at
the Glu2D1 locus. As shown in Table 2 , the quality of
flours from W7268272958 was better in many respects
compared to that from W7268212168 , 50. 6 % higher in
wet gluten content , 47. 8 % higher in Zeleny Value , 3. 5
min longer in development time , 3. 0 min longer in Sta2
bility , and a better Evaluation Value at 2 7 . Therefore ,
Table 1 Relative contents of individual subunits of HMW2GS in various wheat lines
Wheat line
HMW2GS
composition
Relative content of individual subunits
1 7 8 9 2 5 10 12
Chinese Spring N ,7 + 8 ,2 + 12 - 27. 69 15. 98 - 9. 72 - - 42. 86
81224 N ,7 + 9 ,2 + 12 - 28. 50 - 12. 76 18. 18 - - 36. 21
W7268212168 1 ,7 + 9 ,5 + 10 2. 88 23. 62 - 20. 99 - 19. 00 25. 49 -
W7268272958 1 ,7 + 9 ,5 + 12 2. 23 29. 66 - 9. 64 - 16. 22 - 34. 96
W7268242301 1 ,7 + 9 ,5 + 12 3. 43 30. 74 - 9. 21 - 19. 89 - 33. 77
Table 2 Major quality parameters of flours from W7268272958 , its siblines and parental lines (Chengdu , 2000)
Quality parameter Line
W7268272958 W7268212168 81224 Durum779 W7268242301
HMW subunit composition 1 ,7 + 9 ,5 + 12 1 ,7 + 9 ,5 + 10 N ,7 + 9 ,2 + 12 20 1 ,7 + 9 ,5 + 12
Protein content ( %) 16. 01 13. 08 13. 93 14. 98 12. 4
Wet gluten content ( %) 39. 3 26. 1 31. 0 36. 1 -
Zeleny value (mL) 40. 5 27. 4 34. 4 29. 9 -
SDS2sedimentation(mL) 51 43 46 44 52
Water absorption( %) 61. 6 59. 0 61. 4 - -
Development time (min) 5. 0 1. 5 3. 5 - -
Stability(min) 4. 0 1. 0 4. 0 - -
Degree of softening(FU) 90 180 120 - -
Breakdown time (min) 7. 5 2. 0 6. 5 - -
Evaluation value 57 30 46 - -
645 　　　作 　　物 　　学 　　报 30 卷 　

we conclude that the new line containing the 5 + 12
HMW2glutenin subunits had the flours with better baking
quality as compared to those containing the 5 + 10 sub2
units.
3 　Discussion
Using inter2specific crosses (Durum wheat ×bread
wheat) and selecting by SDS2PAGE , We have obtained a
novel stable wheat line with the rare combination of 5 + 12
HMW glutenin subunits. Flours from the line is better
than those from lines contain the 5 + 10 subunit combina2
tion in many baking quality aspects , such as sedimenta2
tion values , wet gluten content , development time and
stability time of farinograph , which is consistent with the
finding of Pena (1995) [4 ] . The further studies on the
baking quality of flours from the line are under way.
The better baking quality associated with the 5 + 12
subunit combination may result from a better synergistic
relationship between the subunits 5 and 12. Analyses of
relative contents of subunits in HMW2GS showed 53. 66 %
of 5 + 12 subunits encoded at Glu2D1 locus , higher than
44. 49 % of 5 + 10 subunits. This is consistent with the
finding by Payne et al . [3 ] that the Glu2D1 locus was the
determinant of baking quality , and that the subunit 12 en2
coded by 1 Dy allele had a higher relative content in lines
containing the 5 + 12 combination than the subunit 10 in
lines having 5 + 10 combination. Shewry et al . also
showed that the 1 Dy subunit was a determinant for the
dimerization of glutenin[10 ] . This might be due to the
higher cysteine content in the y2type subunit , which might
favor the formation of di2sulfide bonds and dimerization of
glutenin as shown by Weegle et al . ( 1996) [11 ] . The
larger amount of y2type subunit might enhance the larger
polymer of glutenin to yield a higher strength of wet gluten
and dough quality. Our data is consistent with these find2
ings.
SDS2PAGE analyses of HMW2GS subunits for choos2
ing the parental lines and the selection of progenies were
the effective , speedy , and with smaller samples. Together
with triple crosses , back crosses and increase of genera2
tion in different growth condition ( such as greenhouse) ,
combining with SDS2PAGE and PCR technology , the 5 +
12 subunit combination could be efficiently and accurately
transfered to wheat lines of high yield and drastically in2
creased the efficiency of breeding.
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745　6 期 WANG Tao et al . : Identification and Characterization of a Novel Wheat Line Containing High Molecular Glutenin Subunits 5 + 12