Introduction
Common wheat (Triticum aestivum L.) is the most widely grown wheat and it is one of the major food crops utilized all over the world. Identification and characterization of the genetic resources of wheat are highly desirable because the available information can be applied in further research and breeding. The improvement of wheat for use of the end-product has become an important part of wheat breeding programmes worldwide. It has been documented that quality is determined principally by the molecular structure of the seed storage proteins, glutenins and gliadins (Bushuk, Reference Bushuk1998). Thus, not only protein quantity but also their quality is of great importance. High-molecular weight-glutenin subunits (HMW-GS) play a major role in this regard. The HMW-GS are encoded by genes at three Glu-1 loci (Glu-A1, Glu-B1 and Glu-D1), which are located on the long arms of homologous group-one chromosomes. These loci encode an x-type subunit of higher molecular weight and a y-type subunit of lower molecular weight. It was demonstrated that good bread-making quality is highly associated with the presence of specific HMW-GS (Payne et al., Reference Payne, Nightigale, Krattinger and Holt1987). Since then, a number of novel GS have been identified and characterized. These novel alleles may be a potential genetic resource for the creation of wheat lines with improved quality (Gálová et al., Reference Gálová, Michalík, Knoblochová and Gregová2002; Dotlačil et al., Reference Dotlačil, Bradová, Hermuth, Stehno, Pogna, Romano, Pogna and Galterio2003; Ren et al., Reference Ren, Wang, Xu, Yang and Ren2008). Databases that contain information on wheat genotypes/cultivars and their HMW-GS compositions may offer breeders the prospect of further advancement by selecting wheat cultivars with beneficial HMW-GS. The objective of our study was to determine the composition of HMW-GS in 120 wheat cultivars (T. aestivum L.).
Materials and methods
A total of 120 hexaploid wheat cultivars (T. aestivum L.), provided by the Gene Bank of the Slovak Republic in Piešťany, were analyzed in this study. The wheat samples originated from 11 countries: Slovakia (SVK, 12), Czech Republic (CZE, 12), France (FRA, 12), Austria (AUT, 12), Italy (ITA, 12), Hungary (HUN, 12), former Yugoslavia (YUG, 12), Ukraine (UKR, 6), Russia (RUS, 6), Great Britain (GBR, 12) and the USA (12). The HMW-GS were extracted from randomly selected single seeds. Protein extraction, electrophoretic separation of glutenins and the detection procedures used were in accordance with the International Seed Testing Association (ISTA) standard procedure for use of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) (Wrigley, Reference Wrigley, Linskens and Jackson1992). HMW-GS were identified by following the catalogue of HMW-GS alleles (Payne and Lawrence, Reference Payne and Lawrence1983). In addition, the presence or absence of the secalin block (1BL.1RS translocation) in the genotypes was examined using acid polyacrylamide gel electrophoresis (A-PAGE), according to the standard ISTA reference method (Draper, Reference Draper1987). The bread-making quality of the grain was expressed as a score (Glu-score and Rye-score) derived from the presence or absence of specific high-molecular weight-glutenins or -gliadins.
Results and discussion
Fourteen alleles and 34 allelic compositions were detected in the set of 120 cultivars. The most frequent HMW-GS patterns were null, 7+9 and 5+10, which were present in 21 cultivars. Ten of the wheat accessions analyzed (8.3%) were observed to be heterogeneous in their glutenin profiles. Five cultivars were heterogeneous at one locus, two cultivars at two loci and another three varieties were heterogeneous at all three Glu-1 loci.
Three alleles were detected at the Glu-A1 locus. The most abundant allele was Glu-A1c, encoding the null subunit (57.1%, Table 1). It was found mostly in cultivars bred in SVK, CZE, YUG and GBR (Table 2). In contrast, the alleles Glu-A1b (subunit 2*) and Glu-A1a (subunit 1) were present in only 14 and 29% of the genotypes, respectively. These alleles were observed mainly in Hungarian, Italian, Austrian and American wheat samples (Table 2). A high frequency of alleles encoding subunits 1 and 2* in wheat cultivars from the USA was observed also by Shan et al. (Reference Shan, Clayshulte, Haley and Byrne2007). These subunits (1, 2*) have a positive impact on the rheological properties of dough, so they are highly desirable in cultivars used for bread-making.
Table 1 The frequencies of HMW-GS coded at the Glu-1 loci
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Table 2 The frequencies (%) of HMW-GS at the Glu-1 loci according to the country of origin
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A total of nine HMW-GS were identified at the Glu-B1 locus. Subunit 7+9 was the predominant one (43.3%, Table 1); it was abundant mostly in wheat bred in HUN, SVK, YUG, RUS and UKR. The subunit 6+8, which is associated with poor bread-making quality, was highly frequent in British and Austrian wheat samples (Table 2). Low-frequency HMW-GS, such as 13+16, 20, 21, 7 and 18, which are encoded by alleles at the Glu-B1 locus were also detected (Table 1). An unpaired subunit 7 was present mainly in French cultivars and also in wheat samples originating from USA, RUS and ITA (Table 2). The rare subunit 21 was found in the cultivar Mironovskaja ulucsennaja (UKR). Subunit 18, which is expressed commonly coupled with subunit 17, was observed as an unpaired subunit in the Italian cultivar Glutinoso. Redaelli et al. (Reference Redaelli, Ng and Pogna1997) observed expression of unpaired subunit 6 in some cultivars of T. aestivum L. They assumed that it was derived from the HMW-GS pair 6+8 (because it had the same mobility as subunit 6 of this pair), but that the expression of only subunit 6 was caused by a switch-off mutation in the coding sequence or in the regulatory region of the gene for subunit 8. Yuan et al. (Reference Yuan, Chen, Zhang, Yan, Zheng and Liu2009) characterized two inactive y-type genes for Glu-B1 in T. aestivum ssp. yunnanese and ssp. tibetanum. They indicated that the silenced 1By genes were 1By9 and that the silencing was caused by deletions of nucleotide A, which resulted in frameshift mutations responsible for the presence of premature stop codons.
Most of the cultivars (61.9%, Table 1) were found to express the subunit pair 5+10, which is encoded by allele Glu-D1d at the Glu-D1 locus. It has been shown that varieties that contain the pair 5+10 can form stronger dough than those that contain subunits 2+12 (Payne, Reference Payne1987; Liang et al., Reference Liang, Tang, Pena, Singh, He, Shen, Yao, Xia and He2010). Thus, wheat carrying the pair 2+12 can be utilized in the production of cookies, crackers or pasta rather than in bread-making. In this study, subunit pair 2+12, encoded by allele Glu-D1a, was observed frequently in cultivars bred in ITA and GBR (Table 2). A high frequency of subunit pair 2+12 has been published for wheat originating in Asian countries, such as China and Japan (Nakamura, Reference Nakamura2001; Li et al., Reference Li, Huang, Sui, Fan, Li and Chu2009), where wheat is used predominately in the production of noodles. Our analysis revealed also the presence of subunit 4+12, encoded by allele c at the Glu-D1 locus, in one cultivar that originated in the USA.
The wheat–rye 1BL.1RS translocation was identified in 25 cultivars by use of A-PAGE. No cultivars that originated in USA and FRA were found to carry this translocation. The presence of the 1BL.1RS translocation has a negative effect on the rheology of dough; it produces a sticky dough with a lack of overmixing, and low loaf volumes (Graybosch, Reference Graybosch2001; Gobaa et al., Reference Gobaa, Brabant, Kleijer and Stamp2008; Zheng et al., Reference Zheng, Byrne, Bai, Shan, Reid, Haley and Seabourn2009).
The Glu-score and Rye-score varied greatly; some samples reached the maximum value of 10 (cultivars GK Kapos, GK David, Vlasta, Baltimor, Luzanovka, Maverick and Smuggler). These cultivars are supposed to have superior bread-making quality. The highest average value of Glu-score was achieved by Hungarian cultivars (8.1). In contrast, the HMW-GS composition of British wheat samples resulted in an average value of only 5.9.
Acknowledgements
This research was supported by the Slovak Research and Development Agency under the contracts APVV LPP-025107 and VMSP-P-0055-09.