Molecular detection of high- and low-molecular-weight glutenin subunit genes in common wheat cultivars from 20 countries using allele-specific markers

2011 ◽  
Vol 62 (9) ◽  
pp. 746 ◽  
Author(s):  
H. Jin ◽  
J. Yan ◽  
R. J. Peña ◽  
X. C. Xia ◽  
A. Morgounov ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecylsulfate ◽  
Functional Markers ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
High Frequencies ◽  
Allele Specific ◽  
End Use

The composition and quantity of high- and low-molecular-weight glutenin subunits (HMW-GS and LMW-GS) plays an important role in determining the end-use quality of wheat products. In the present study, 718 wheat cultivars and advanced lines from 20 countries were characterised for the HMW-GS and LMW-GS with allele-specific molecular markers. For the Glu-A1 locus, 311 cultivars (43.3%) had the subunit Ax2*, which predominated in cultivars from Canada (83.3%), Romania (91.7%), Russia (72.2%) and USA (72.2%). At Glu-B1 locus, 197 cultivars (27.4%) contained the By8 subunit and its frequency was higher in Japanese (60.0%) and Romanian (62.5%) genotypes than in those from other countries; 264 cultivars (36.8%) carried the By9 subunit, mostly existing in the cultivars from Austria (100.0%), Russia (72.2%), and Serbia (72.7%); the By16 subunit was present in 44 cultivars (6.1%), with a relatively high percentage in Chile (19.5%), whereas almost no cultivars from other countries had this subunit; the frequency of Bx7OE was 3.1%, and was found only in cultivars from Argentina (12.1%), Australia (4.1%), Canada (25.0%), Iran (20.0%), and Japan (30.0%). There were 446 genotypes (62.1%) with the subunit Dx5 at the Glu-D1 locus; high frequencies of Dx5 occurred in cultivars from Hungary (90.0%), Romania (95.8%), and Ukraine (92.3%). At the Glu-A3 locus, the frequencies of Glu-A3a, b, c, d, e, f and g were 2.9, 6.8, 53.2, 12.8, 7.7, 13.8, and 2.4%, respectively. Glu-A3a was detected only in the cultivars from Bulgaria (13.3%), China (12.2%), Germany (2.7%), Iran (6.7%), Mexico (14.3%), Turkey (4.7%), and USA (5.1%); the high frequencies of superior alleles Glu-A3b and d were found in cultivars from Australia (39.7%) and France (24.5%); Glu-A3c was widely distributed in cultivars from all the countries; the high frequencies of Glu-A3e, f and g were detected in cultivars from Argentina (33.3%), Canada (29.2%), and Hungary (20.0%). At the Glu-B3 locus, Glu-B3a, b, c, d, e, f, g, h and i were present in frequencies of 0.4, 22.3, 0.3, 2.8, 1.9, 3.9, 27.2, 18.8, and 7.1%, respectively. Glu-B3a was detected only in cultivars from Argentina (3.0%) and Ukraine (15.4%) cultivars; high frequencies of Glu-B3b and d were found in the cultivars from Romania (62.5%) and Mexico (14.3%); Glu-B3c was detected only in Romanian (8.3%) genotypes; frequencies of e, f, h and i were high in cultivars from Austria (40.0%), China (14.3%), USA (43.0%), and Argentina (33.3%); Glu-B3g was mostly detected in the cultivars from Germany (69.3%), Norway (77.3%), and Serbia (63.6%). The frequency of the 1B·1R translocation was 13.4%; it occurred in cultivars from all the countries except Australia, Austria, Norway, and Serbia. The functional markers applied in this study, in agreement with the results of sodium-dodecylsulfate–polyacrylamide gel electrophoresis, were accurate and stable, and can be used effectively in wheat quality breeding.

Characterisation of high- and low-molecular-weight glutenin subunit genes in Chinese winter wheat cultivars and advanced lines using allele-specific markers and SDS-PAGE

2010 ◽  
Vol 61 (1) ◽  
pp. 84 ◽  
Author(s):  
F. P. Yang ◽  
L. H. Wang ◽  
J. W. Wang ◽  
X. Y. He ◽  
X. K. Zhang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Molecular Markers ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
Specific Pcr ◽  
Sds Page ◽  
Allele Specific ◽  
Chinese Wheat

Wheat end-use product quality is highly influenced by the composition and quantity of high- and low-molecular-weight glutenin subunits (HMW-GS and LMW-GS). In the present study, 224 Chinese wheat cultivars and advanced lines were characterised for the HMW-GS and LMW-GS with allele-specific PCR markers and sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that 56 cultivars (25.0%) carried the allele Glu-D1-1d (Dx5), while 80 cultivars (35.7%) with the allele Glu-B1-2a (By8) produced a 527-bp specific band. Fourteen genotypes (6.3%) with the allele Glu-B1e (Bx20) yielded a 701-bp amplicon with the marker Mar and a 753-bp specific PCR fragment with the marker ZSBy9aF1/R3. Glu-B1h (Bx14+By15) was present in only 1 genotype, and 2 cultivars contained the allele Glu-B1f (Bx13+By16) identified with the marker ZSBy9F2/R2. Four genotypes (1.8%) with the allele Glu-B1-1d (Bx6) gave 695-bp and 830-bp bands, and 5 genotypes (2.2%) with the allele Glu-B1i (Bx17+By18) amplified a 659-bp fragment using the marker Bx. One hundred and six cultivars (47.3%) had the allele Glu-B1-2b (By9), amplifying a 663-bp fragment with the marker ZSBy9aF1/R3; 34 genotypes (15.8%) contained the allele Glu-B3d, generating a 662-bp PCR fragment with the marker gluB3d. Fifteen cultivars (7.0%) with the allele Glu-B3b yielded 1570-bp and 750-bp PCR amplicons with the markers gluB3b and gluB3bef, respectively. The allele Glu-B3h was found in 7 cultivars, generating a 1022-bp PCR fragment with the marker gluB3h. The genotypes detected by SDS-PAGE were mostly consistent with those identified by molecular markers, confirming the utility of the molecular markers. The information for the HMW-GS and LMW-GS in Chinese wheat cultivars will be useful in wheat breeding programs.

IDENTIFICATION OF CANADIAN AND AMERICAN WHEAT CULTIVARS BY SDS GRADIENT PAGE ANALYSIS OF GLIADIN AND GLUTENIN SUBUNITS

1987 ◽  
Vol 67 (4) ◽  
pp. 945-952 ◽  
Author(s):  
B. A. MARCHYLO
Keyword(s):  
Molecular Weight ◽  
Spring Wheat ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
Glutenin Subunits ◽  
Molecular Weights ◽  
Additional Protein ◽  
Hmw Glutenin

Sodium dodecyl sulphate gradient polyacrylamide gel electrophoresis (SDSGPAGE) was used to resolve gliadin and high- and low-molecular-weight glutenin subunits from 19 registered Canadian spring wheat cultivars eligible for Canada Western Red Spring (CWRS) and Canada Prairie Spring (CPS) wheat grades and eight nonregistered spring wheat cultivars from the U.S.A. Reproducible molecular weight estimates were obtained for wheat proteins of apparent molecular weights ranging from 34 238 to 136 174 (avg. CV = 0.72%). Eight different patterns of HMW glutenin subunits consisting of 7–11 protein bands were observed for the 27 cultivars and their biotypes. SDSGPAGE was able to discriminate among the majority of cultivars with all non-registered cultivars and their biotypes distinguishable from registered cultivars. Separation of glutenin subunits along with gliadins provided additional protein bands which assisted in the discrimination of cultivars.Key words: SDS gradient PAGE, wheat cultivar identification, gliadin, glutenin subunits

Rapid Analytical Techniques For Measuring The High And Low Molecular Weight Contents Of Urokinase Samples

1981 ◽  
Author(s):  
E M Devine ◽  
L L Shen ◽  
A C Saldivar ◽  
R L Gendrich ◽  
R G Wiegand ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Molecular Species ◽  
Biological Activities ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecylsulfate ◽  
Gel Filtration ◽  
Molecular Forms ◽  
Low Molecular Weight ◽  
Fibrin Plate ◽  
Filtration Technique

Two assay procedures have been developed for measuring the ratio of the two molecular weight forms of urokinase commonly found in either urine or the fluid of cultured human kidney cells. The first method is a mini gel filtration technique which rapidly separates the high molecular weight (HMW) and low molecular weight (LMW) urokinases. This procedure can be used for urokinase samples of any purity. The biological active fractions are assayed by the fibrin plate technique. These biological activities are converted to relative molar amounts by the use of the molar activity constant for each molecular weight form. The second procedure is based on the differing responses of the two molecular species in the fibrin plate and the Ploug clot lysis assay systems. The comparison of assay responses of an unknown with a predetermined curve obtained from known mixtures of HMW and LMW urokinases results in a measure of the relative amounts of the two molecular forms. This system is not applicable to samples of low purity.The two systems gave comparable results. For example, the WHO First International Reference Preparation (66/46) of Urokinase is 31% HMW urokinase by the differential method and 32% HMW urokinase by the gel filtration technique. Both methods require small amounts of sample (400 IU or less) and should replace the use of sodium dodecylsulfate polyacrylamide gel electrophoresis for urokinase molecular species determinations.

Transient changes in wheat endosperm polymeric proteins during the later stages of grain development

1995 ◽  
Vol 75 (1) ◽  
pp. 195-198 ◽  
Author(s):  
A. Hussain ◽  
O. M. Lukow
Keyword(s):  
Molecular Weight ◽  
Storage Proteins ◽  
Developmental Changes ◽  
Wheat Cultivars ◽  
Grain Development ◽  
Gluten Proteins ◽  
Distal Half ◽  
End Use ◽  
Qualitative Changes

End-use quality of wheat depends mainly on the composition of the storage proteins. These proteins undergo changes during grain development. This study was carried out to examine modifications in the polymeric protein fraction at different days post anthesis (DPA) and to determine whether these changes are cultivar dependent. Endosperm proteins of three hexaploid wheat cultivars harvested at different stages of maturity and extracted sequentially from distal half kernels showed quantitative and qualitative changes when separated in a 7.5–9.0% gradient polyacrylamide gel system. Sudden qualitative changes were observed at a later stage of seed development (35 DPA) in all tested cultivars. Most qualitative changes occurred in proteins of molecular weight 60–90 kDa and least in proteins above 90 kDa. An intense 60-kDa band appeared in the 35-DPA sample of Columbus; this band faded as the grain approached full maturity (40 DPA). Key words: Developmental changes, electrophoresis, gluten proteins, seed ripening

scholarly journals Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars

2021 ◽  
Vol 22 (14) ◽  
pp. 7709
Author(s):  
Kyoungwon Cho ◽  
You-Ran Jang ◽  
Sun-Hyung Lim ◽  
Susan B. Altenbach ◽  
Yong Q. Gu ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Allelic Variation ◽  
Reversed Phase ◽  
Glutenin Subunit ◽  
Low Molecular Weight ◽  
Near Isogenic Lines ◽  
Wheat Cultivars ◽  
Reliable Determination ◽  
Isogenic Lines

The low-molecular weight glutenin subunit (LMW-GS) composition of wheat (Triticum aestivum) flour has important effects on end-use quality. However, assessing the contributions of each LMW-GS to flour quality remains challenging because of the complex LMW-GS composition and allelic variation among wheat cultivars. Therefore, accurate and reliable determination of LMW-GS alleles in germplasm remains an important challenge for wheat breeding. In this study, we used an optimized reversed-phase HPLC method and proteomics approach comprising 2-D gels coupled with liquid chromatography–tandem mass spectrometry (MS/MS) to discriminate individual LMW-GSs corresponding to alleles encoded by the Glu-A3, Glu-B3, and Glu-D3 loci in the ‘Aroona’ cultivar and 12 ‘Aroona’ near-isogenic lines (ARILs), which contain unique LMW-GS alleles in the same genetic background. The LMW-GS separation patterns for ‘Aroona’ and ARILs on chromatograms and 2-D gels were consistent with those from a set of 10 standard wheat cultivars for Glu-3. Furthermore, 12 previously uncharacterized spots in ‘Aroona’ and ARILs were excised from 2-D gels, digested with chymotrypsin, and subjected to MS/MS. We identified their gene haplotypes and created a 2-D gel map of LMW-GS alleles in the germplasm for breeding and screening for desirable LMW-GS alleles for wheat quality improvement.

scholarly journals Thermolability of 28S ribosomal ribonucleic acid from the liver of Crotalus durissus terrificus (Ophidia, Reptilia)

1973 ◽  
Vol 135 (1) ◽  
pp. 73-79 ◽  
Author(s):  
J. F. Giorgini ◽  
F. L. De Lucca
Keyword(s):  
Molecular Weight ◽  
Gradient Centrifugation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sucrose Density Gradient ◽  
Dimethyl Sulphoxide ◽  
Low Molecular Weight ◽  
28S Rrna ◽  
Sucrose Density Gradient Centrifugation ◽  
Crotalus Durissus Terrificus ◽  
Crotalus Durissus

Instability of 28S rRNA of Crotalus durissus terrificus liver was observed during hotphenol extraction: purified 28S rRNA is converted into an 18S RNA component by heat treatment. It was also found that ‘6S’ and ‘8S’ low-molecular-weight RNA species were released during the thermal conversion. This conversion and the release of the low-molecular-weight species were also induced by 8m-urea and 80% (v/v) dimethyl sulphoxide at 0°C. Evidence is presented that this phenomenon is an irreversible process and results from the rupture of hydrogen bonds. The 18S RNA product was shown to be homogeneous by polyacrylamide-gel electrophoresis and by sucrose-density-gradient centrifugation. The base composition of the 18S RNA products obtained by heat, urea or dimethyl sulphoxide treatments was similar. The C+G content of the 18S RNA product was different from that of the native 18S rRNA, but similar to that of 28S rRNA.

Normal Mature Human Enamel Protein

1979 ◽  
Vol 58 (2_suppl) ◽  
pp. 986-987 ◽  
Author(s):  
A. Belcourt
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Organic Matrix ◽  
Low Molecular Weight ◽  
Human Enamel ◽  
Weak Density ◽  
Enamel Protein

Pure enamel was prepared using an original microdissection technic. Protein concentration was 375 μg per gram of enamel. Polyacrylamide gel electrophoresis showed a single fast-migrating zone containing a thin double band. Ultracentrifugation studies suggested that the proteins were of low molecular weight or of weak density. Absorption spectra showed a strong absorbance at 260nm. Amino acid analyses yielded a composition of 25% Gly, 13.5% Glu, 11% Ser, 11% Pro, 2% Cys and 2% Hyp. A glucidic content of 15% was estimated and glucose, galactose, mannose and fucose were identified. The organic matrix of enamel seemed to be constituted of two major glycoproteins probably fibrous but different from keratin.

scholarly journals Differential induction of allergy responses by low molecular weight wheat proteins from six wheat cultivars

2017 ◽  
Vol 60 (1) ◽  
pp. 55-59
Author(s):  
Miju Cho ◽  
Hyeri Lee ◽  
Min Hee Hwang ◽  
Young-Keun Cheong ◽  
Chon-Sik Kang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
Wheat Proteins ◽  
Differential Induction
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