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Wheat Information
Service
Number 76: 77-79 (1993)
Detection
of HMW glutenin genes by DNA hybridization and breadbaking quality of
amphidiploid synthesized between Aegilops squarrosa and
Secale cereale.
S. Sekiguchi1, J. Ono1, and T.
Taira2
1 Central Laboratory, Nippon Flour Mills Co. Ltd.,
Atsugi, Kanagawa
2
Lab. of Genetics and
Plant Breeding, College of Agriculture, Univ. of Osaka Prefecture,
Sakai, Osaka 593, Japan
We have synthesized a new amphidiploid having genomes DDRR between
Aegilops squarrosa var. typica and Secale cereale
cv. Prolific (Kawakubo and Taira 1992). High molecular
weight (HMW) glutenin subunits are governed by genes located on long
arms of homoeologous group 1 chromosomes in hexaploid wheat (Boyd et
al 1967, Payne et al 1979) and all three HMW glutenin loci are
Glu-A1, Glu-B1, and Glu-D1 (Payne and Lawrence 1983).
Since alleles at these three loci have shown differential effects on
bread-baking quality in hexaploid wheat, the structure of HMW
glutenin genes of genomes D and R, and the flour quality of
amphidiploid were investigated in this study.
Identification and comparison of genomic DNA restriction-fragment
of HMW glutenin genes between amphidiploid and its direct parents
Amino acid sequence of protein encoded by a HMW glutenin cDNA
fragment (777bps) isolated from hexaploid wheat cv. Park consisted of
repetitive units of two motif, PGQGQQ and GYYPTSLQQ, which were
typical to the amino acid sequence of common region in HMW glutenin
proteins. Enzyme-digested genomic DNA fragments from F3
progeny of amphidiploid and its direct parents, tetraploid Ae.
squarrosa (genomes DDDD) and S. cereale (RRRR),
were probed with the cDNA fragment. The hybridization patterns of DNA
restriction fragments were as follows (Fig.
1): in
EcoRI digestion, five bands were found in amphidiploid, two
bands in Ae. squarrosa and three bands in S.
cereale; in BamHI digestion, five bands were
found in amphidiploid, two bands in Ae. squarrosa and three
bands in S. cereale; in HindIII
digestion, five bands were found in amphidiploid, two
bands in Ae. squarrosa and four bands in S. cereale.
In particular, the band c in EcoRI digestion of S.
cereale (Fig.
1B, lane 8) had
slightly large molecular weight than the band a of amphidiploid (lane
7), and the band d (lane 8) was the same as the band a. It seems that
the band c in S. cereale splits into two fragments such
as bands a and b in amphidiploid.
Comparison of HMW glutenin proteins of amphidiploid and its
parents
Glutenin proteins deposited in endosperm were extracted and
separated by SDS PAGE (Fig.
2). The
distribution of HMW glutenin proteins in diploid seeds was different
from that in tetraploid seeds in both parental lines. Origin of
protein bands of amphidiploid was considered as follows: band a was
derived from either S. cereale or Ae. squarrosa;
bands b, c, e and f from S. cereale; band d from
Ae. squarrosa.
Baking tests
Bread was baked with 30 g of flour by straight dough procedure.
Because of bad weather condition at harvesting time, amphidiploid
flour was low quality, and thus the dough was sticky and runny.
Nevertheless, a loaf of amphidiploid was increased by 29% in volume
compared with that of rye, although amphidiploid showed 49% in volume
of bread wheat (Fig.
3). Wet gluten
(3%) was obtained from amphidiploid flour, but no gluten was detected
in rye flour.
These results showed that the amphidiploid in which Glu-1D
gene of glutenin subunits was introduced from Ae. squarrosa
had improved bread-baking quality in comparison with rye.
References
Boyd WJR, Lee JW, and Wrigley CW (1967), The control of
wheat gluten synthesis at the genome and chromosome level.
Experientia 23: 332-333.
Kawakubo J and Taira T (1992) Intergeneric hybrids between
Aegilops squarrosa and Secale cereale and their meiotic
chromosome behavior. Plant Breeding 109: 108-115.
Payne Pl, Law CN, and Mudd EE (1979) Control by homoeologous group 1
chromosome of the high molecular weight subunits of glutenin: A major
protein of wheat endosperm. Theor. Appl. Genet. 58: 113-120.
Payne PI, and Lawrence GJ (1983) Catalogue of alleles for the complex
gene loci, Glu-A1, Glu-B1 and Glu-D1 which code for
high molecular weight subunits of glutenin in hexaploid wheat. Cereal
Res. Commun. 11: 29-35.
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