Development of simple and co-dominant PCR markers to genotype puroindoline a and b alleles for grain hardness in bread wheat (Triticum aestivum L.)

Grain hardness is one of the most important quality characteristics of cultivated bread wheat (Triticum aestivum L.). A large deletion in the puroindoline a (Pina) gene or single nucleotide polymorphisms (SNPs) in the puroindoline b (Pinb) gene results in hard grain texture. So far, nine Pina alleles (Pina-D1a–Pina-D1b, Pina-D1k–Pina-D1q) and seventeen Pinb alleles (Pinb-D1a–Pinb-D1g, Pinb-D1p–Pinb-D1ab) have been identified in bread wheat. The major Pina and Pinb alleles identified in hard wheat cultivars are Pina-D1b, Pinb-D1b, Pinb-D1c and Pinb-D1d. In this study, a three-primer PCR system was employed to develop nine co-dominant STS markers for genotyping Pina-D1a and Pina-D1b, whereas temperature-switch (TS) PCR was used to develop six co-dominant SNP markers for genotyping the Pinb-D1a, Pinb-D1b, Pinb-D1c and Pinb-D1d alleles. These STS and TS-PCR markers were used to verify the grain hardness genotype of 100 wheat cultivars. The reliability and genotyping accuracy of TS-PCR markers were confirmed through sequencing of PCR products and a comparison with previously published results. Therefore, STS and TS-PCR markers offer a simple, cost-effective and reliable method for high-throughput genotyping Pina and Pinb alleles to select grain hardness in wheat quality breeding programs and for wheat market classification.     

Effects of allelic variation of HMW-GS and LMW-GS on mixograph properties and Chinese noodle and steamed bread qualities in a set of Aroona near-isogenic wheat lines

High-molecular-weight glutenin (HMW-GS) and low-molecular-weight glutenin (LMW-GS) subunits play an important role in determining wheat quality. To clarify the contribution of each subunit/allele to processing quality, 25 near-isogenic lines with different HMW-GS and LMW-GS compositions grown at two locations during the 2010 cropping season were used to investigate the effects of allelic variation on milling parameters, mixograph properties, raw white Chinese noodle (RWCN) and northern style Chinese steamed bread (NSCSB) qualities. The results showed that Glu-B1 and Glu-B3 made a large contribution to determining mixograph properties and processing quality, respectively. Subunit pairs 17 + 18 and 5 + 10, and alleles Glu-A3b, Glu-A3d, Glu-B3g and Glu-D3f made significant contributions to mixograph properties and no significant difference was detected on most parameters of RWCN and NSCSB for the allelic variation of HMW-GS and LMW-GS. The allelic interactions among glutenin loci had significant effects on wheat quality. The line with 1, 17 + 18, 2 + 12, Glu-A3c, Glu-B3b, Glu-D3c associated with superior mixograph properties, the line with 1, 7 + 9, 2 + 12, Glu-A3c, Glu-B3d, Glu-D3c had superior viscoelasticity of RWCN, and the line with 1, 7 + 9, 2 + 12, Glu-A3e, Glu-B3b, Glu-D3c had the highest total score of NSCSB. These results provide useful information for genetic improvement of the qualities of traditional Chinese wheat products.     

中国小麦微核心种质 Glu-A3位点等位变异的分布

低分子量麦谷蛋白亚基在面包和面条食品加工过程中起着十分重要的作用。为了便于对含有LMW-GS优良基因的亲本筛选,选取来源于不同麦区包括地方品种和育成品种在内的208份核心种质为供试材料,用特异PCR方法检测Glu-A3位点LMW-GS基因的等位变异。结果表明,Glu-A3d出现频率(26.4%)明显高于其他等位类型,在地方品种和育成品种中的分布频率分别为26.7%和26.0%;分布频率从大到小依次为Glu-A3d>Glu-A3c>Glu-A3a>Glu-A3b>Glu-A3e>Glu-A3g>Glu-A3f;在来源于冬春兼播麦区的育成品种中未检测出Glu-A3e和Glu-A3f。对面筋强度贡献较大的Glu-A3b在地方品种和育成品种中的分布频率分别为10.7%和18.2%,说明我国小麦总体品质水平有了较大的提升。     

Novel allelic variants encoded at the Glu-D3 locus in bread wheat

Low-molecular weight glutenin subunits (LWM-GS) are important components of wheat (Triticum aestivum L.) gluten, with important effects on end-use quality. The LMW-GS are encoded at Glu-3 loci (Glu-A3, Glu-B3 and Glu-D3, on the short arms of chromosomes 1A, 1B and 1D), each of which exhibits extensive allelic variation. Each locus encodes numerous LMW-GS, some of which have similar electrophoretic mobilities, making it difficult to distinguish among Glu-3 loci. Alleles of the Glu-D3 locus of bread wheat are considered the most problematic to assign. To date, six Glu-D3 alleles, designated a, b, c, d, e and f, have been reported. We report five previously undescribed alleles (g, h, i, j and k), and describe a method for characterizing them using a combination of SDS-PAGE and multiplexed PCR-based DNA markers. This method could be used for accurate identification of Glu-D3 alleles, permitting the estimation of the effects of these alleles on end-use quality and the selection of desirable alleles and allelic combinations in wheat breeding.     

Comparative analyses of LMW glutenin alleles in bread wheat using allele-specific PCR and SDS-PAGE

One hundred and eighty-two bread wheat cultivars developed in India were characterized for low molecular weight (LMW) glutenins using SDS-PAGE and allele-specific polymerase chain reaction (PCR) to assess allelic diversity encoded by Glu-3 loci, as well as their utility for correctly identifying different alleles. SDS-PAGE indicated Glu-A3c is present in 64.6% of the cultivars, Glu-A3b in 13.8%, Glu-A3d in 12.7% and Glu-A3e/f in 8.8%. Seven types of alleles were present at the Glu-B3 locus: Glu-B3b (29.3%), Glu-B3g (27.0%), Glu-B3h (13.8%), Glu-B3i (16.1%), Glu-B3j (12.1%), Glu-B3c (0.6%) and Glu-B3d (1.1%). SDS-PAGE found three types of Glu-D3 alleles: Glu-D3a (30.2%), Glu-D3b (67.1%) and Glu-D3c (2.7%). However, PCR found two different alleles in cultivars classified as carrying Glu-D3a and three alleles in those identified as carrying Glu-D3b cultivars, indicating a more complex nature of the Glu-D3 locus. In conclusion, the data found greater consistency between the SDS-PAGE and PCR amplification patterns of alleles such as Glu-A3c, Glu-A3d, Glu-B3g, Glu-B3h and Glu-B3i, and less consistency between those same patterns in the Glu-A3b, Glu-A3e/f and Glu-B3b alleles. More studies are needed in order to achieve unambiguous identification of the Glu-3 alleles and thereby allow their greater utility in germplasm evaluation and breeding.     

Influence of allelic prolamin variation and localities on durum wheat quality

Thirty-seven varieties of a Mediterranean durum wheat collection grown in Tunisia and Spain were analysed for their allelic composition in prolamins, as well as their protein concentration, sodium dodecyl sulphate sedimentation (SDSS) test and mixograph parameters. Genotype was a greater source of variation in all measurements than locality. Uncommon high and low molecular glutenin subunits (HMW-GS and LMW-GS) were found (V and 2•• subunits at Glu-A1, 13 + 16 at Glu-B1, 5* subunit and ax allele at Glu-A3). The rare combinations 2 + 4+14 + 18 and 8 + 9+13 + 16+18 subunits at the Glu-B3 locus were found. Glu-A3ax had a positive influence on SDSS and mixograph parameters. Of all the prolamins, those that have the B-LMW-GS composition aaa (for Glu-A3, Glu-B3 and Glu-B2 loci, respectively), when associated with the Glu-A1c and Glu-B1d gave the best semolina quality. By contrast, semolina quality is poor when this same composition is associated with the Glu-A1c and Glu-B1e and even poorer when associated with the Glu-A1c and Glu-B1f. In addition, the cultivars with B-LMW-GS allelic composition aab (for Glu-A3, Glu-B3 and Glu-B2 loci, respectively), when associated with the Glu-A1c and Glu-B1d, gave high quality, whereas when associated with the Glu-A1c and Glu-B1e or with Glu-A1o and Glu-B1f, the quality was very poor.     

Definition of the low molecular weight glutenin subunit gene family members in a set of standard bread wheat (Triticum aestivum L.) varieties

Low-molecular-weight glutenin subunits (LMW-GS) are a class of seed storage proteins that play a major role in the determination of the viscoelastic properties of wheat dough. The LMW-GSs are encoded by multi-gene families at the Glu-A3, Glu-B3 and Glu-D3 loci, with more than 15 genes present in most bread wheat varieties. However, the genic profile associated with different alleles has not been clearly defined. Here, the LMW-GSs in a set of standard varieties were analyzed using molecular markers. In most cases, each Glu-3 allele was represented by a specific haplotype; however, some alleles were undistinguishable. The Glu-A3e and Glu-A3g alleles showed an identical marker haplotype, as did the alleles Glu-B3c and Glu-B3d, and Glu-B3f and Glu-B3ab. In contrast, two haplotypes among varieties designated Glu-D3c were differentiated. The marker profiles present at the Glu-D3 locus exhibited less variation compared to the genes at the Glu-A3 and Glu-B3 loci. Results show the potential of the LMW-GS gene marker system in the characterization of the LMW-GS alleles present in specific bread wheat varieties, and its reconciliation with protein-based nomenclature. This approach will advance the understanding of the contribution of each of the LMW-GS gene alleles in the control of the end-use quality.     

小麦高、低分子量麦谷蛋白亚基对品质性状的影响

为给小麦品质改良提供理论依据,以6个国家的532个品种(系)为材料,分析了小麦高、低分子量麦谷蛋白亚基与品质性状的关系。结果表明,各位点上不同的等位变异对品质性状的效应存在显著差异,亚基1、2*、17+18、5+10、GluA3f、GluB3b和GluB3g对面团形成时间和稳定时间具有显著的正向效应;亚基N、1、14+15、17+18、7+8、2+12、3+12、4+12、GluB3d、GluB3f、GluB3h和GluB3g对蛋白质和湿面筋含量具有显著正向效应;从亚基对品质性状的综合影响来看,1、17+18、GluA3f、GluB3g和GluB3f可作为优势亚基。具有1、17+18、2+12和1、17+18、5+10高分子量麦谷蛋白亚基组合的品种(系)综合品质性状显著优于含有N、14+15、2+12亚基组合的品种(系);具有低分子量麦谷蛋白亚基组合GluA3c、GluB3g,GluA3d、GluB3g和GluA3f、GluB3b品种(系)的综合品质性状显著优于含有GluA3b、GluB3j亚基组合的品种(系)。     

High frequency of abnormal high molecular weight glutenin alleles in Chinese wheat landraces of the Yangtze-River region

A total of 485 common landraces of bread wheat were collected from the Yangtze-River region of China. Their high molecular weight glutenin subunit (HMW-GS) composition was analyzed by Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry (MALDI-TOF-MS). Among all landraces tested, 453 were homogeneous for HMW-GS, 32 were heterogeneous, and 37 contained abnormal subunits. A total of 22 alleles were detected, including 3 at Glu-A1, 13 at Glu-B1 and 6 at Glu-D1, respectively. Higher variations occurred at the Glu-B1 locus compared with Glu-A1 and Glu-D1. Glu-A1c (74.0%), Glu-B1b (40.4%), Glu-D1a (84.9%) appeared to be the most frequent alleles at Glu-A1, Glu-B1 and Glu-D1, respectively. Two alleles ("null" and 1) at the Glu-A1 locus, three allele compositions (7 + 8, 7OE + 8, 7 + 9) at the Glu-B1 locus, and two (2 + 12 and 5 + 10) at the Glu-D1 locus appeared to be the common types in the 485 landraces. Sixteen new alleles represented by abnormal subunits were identified at the Glu-B1 and the Glu-D1 locus.     

陕西小麦 Glu A3和 Glu B3位点等位变异的检测和分析

低分子量谷蛋白亚基（LMWGS）与小麦品质密切相关。为了给陕西小麦的品质改良提供参考依据,采用STS分子标记,检测了175份陕西小麦品种（系） GluA3和 GluB3位点的等位变异组成。结果表明,陕西小麦 GluA3位点存在4种等位变异,即 GluA3a、 GluA3b、 GluA3c和 GluA3d,分别占12.6%、1.7%、58.3%和27.4%; GluB3位点存在8种等位变异,即 GluB3a、 GluB3b、 GluB3d、 GluB3e、 GluB3f、 GluB3g、 GluB3i和 GluB3j,分别占4.6%、2.9%、45.7%、0.6%、2.9%、8.5%、4.0%和30.8%。在陕西不同地区小麦之间,两个位点等位变异的种类、组合及其分布比例存在差异,这可能与地区间不同的自然地理环境、饮食习惯、育种目标及亲本选择有关。     

Reduced height alleles (Rht) and Hagberg falling number of wheat

Near-isogenic lines varying for alleles for reduced height (Rht) and photoperiod insensitivity (Ppd-D1) in cv. Mercia (2005/6–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht8c+Ppd-D1a, Rht-D1c, Rht12) and cvs Maris Huntsman and Maris Widgeon (2007/8–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-B1b+Rht-D1b, Rht-D1b+Rht-B1c) were compared at one field site, but within different systems (‘organic’, O, 2005/6–2007/8 v. ‘intensive’, I, 2005/6–2010/11). Further experiments at the site (2006/7–2008/9) compared 64 lines of a doubled-haploid (DH) population [Savannah (Rht-D1b) × Renesansa (Rht-8c+Ppd-D1a)]. Gibberellin (GA) insensitive dwarfing alleles (Rht-B1b; Rht-B1c; Rht-D1b; Rht-D1c) could reduce α-amylase activity and/or increase Hagberg falling number (HFN) but effects depended greatly on system, background and season. Only Rht-B1c increased grain dormancy despite producing plants taller than Rht-D1c. The GA-sensitive Rht8c+Ppd-D1a in Mercia was associated with reduced HFN but analysis of the DH population suggested this was more closely linked with Ppd-D1a, rather than Rht8c. The GA-sensitive severe-dwarfing allele Rht12 was associated with reduced HFN. Instability in HFN over season tended to increase with degree of dwarfing. There was a negative association between mean grain weight and HFN that was in addition to effects of Rht and Ppd-D1 allele.     

Effect of allelic variation at the Glu-3/Gli-1 loci on breadmaking quality parameters in hexaploid wheat (Triticum aestivum L.)

Low molecular weight glutenin subunits (LMW-GS) encoded by the Glu-3 loci are known to contribute to wheat breadmaking quality. However, the specific effect of individual Glu-3 alleles is not well understood due to their complex protein banding patterns in SDS-PAGE and tight linkage with gliadins at the Gli-1 locus. Using DNA markers and a backcross program, we developed a set of nine near isogenic lines (NILs) including different Glu-A3/Gli-A1 or Glu-B3/Gli-B1 alleles in the genetic background of the Argentine variety ProINTA Imperial. The nine NILs and the control were evaluated in three different field trials in Argentina. Significant genotype-by-environment interactions were detected for most quality parameters indicating that the effects of the Glu-3/Gli-1 alleles are modulated by environmental differences. None of the NILs showed differences in total flour protein content, but relative changes in the abundance of particular classes of proteins cannot be ruled out. On average, the Glu-A3f, Glu-B3b, Glu-B3g and Glu-B3i_Man alleles were associated with the highest values in gluten strength-related parameters, while Glu-A3e, Glu-B3a and Glu-B3i_Chu were consistently associated with weak gluten and low quality values. The value of different Glu-3/Gli-1 allele combinations to improve breadmaking quality is discussed.     

新疆小麦品种 Glu A3和 Glu B3位点等位变异的分布

为给新疆小麦品质育种提供理论依据,利用 GluA3、 GluB3位点上的17个STS标记检测了185份新疆冬、春小麦品种 GluA3和 GluB3位点的等位变异。结果表明,新疆小麦品种以 GluA3c、 GluB3a和 GluB3j亚基为主,其分布频率分别为64.86%、22.70%和17.84%。新疆冬、春小麦品种在 GluA3位点上均以 GluA3c亚基为主,分布频率分别为63.30%和67.11%;在 GluB3位点上,新疆冬、春小麦品种分别以 GluB3j和 GluB3a为主,分布频率分别为22.02%和26.32%。新疆冬、春小麦农家品种亚基类型较少,冬小麦农家品种仅有5种类型（以 GluA3c和 GluB3i为主）,春小麦农家品种有10种类型（以 GluA3c和 GluB3d为主）。引进品种和自育品种亚基类型丰富,冬小麦引进品种以 GluA3c和 GluB3i为主,分布频率为12.84%和6.42%;春小麦引进品种以 GluA3c和 GluB3j为主,分布频率为17.11%和6.58%。冬小麦自育品种以 GluA3c和 GluB3j亚基类型为主,分布频率为45.87%和18.35%;春小麦自育品种以 GluA3c和 GluB3a亚基类型为主,分布频率为36.84%和18.42%。     

花药培养与麦谷蛋白亚基分子标记结合选育小麦新品种的研究

为快速获得携带麦谷蛋白优质亚基基因的小麦新品种,提高小麦的品质育种技术水平,利用引进的矮败材料与和尚头、甘春20号、临麦34号等10个不同品种(系)杂交,并对杂交后代进行了花药培养,获得了115份花培株系;利用PCR对花培后代株系及杂交亲本进行了优质贮藏蛋白亚基分子标记检测,3个HMW-GS为 Bx7、 Bx14、 Dx5,3个LMW-GS为 Glu-A3ac、 Glu-A3d、 Glu-B3b。结果表明,在115份花培材料中, Bx7的出现频率最高,为94.78%,其余依次为 Glu-A3ac、 Dx5、 Bx14、 Glu-A3d和 Glu-B3b;获得了44份聚合4个亚基以上的材料;结合农艺性状鉴定,筛选出了3份综合性状优异的小麦新品系AB158、AB167和AB332。本研究将花培育种技术、分子标记辅助选择技术及矮败小麦育种技术进行了有机结合,其结果可为提升小麦品质育种技术水平提供参考。     

Isolation and characterization of EMS-induced Dy10 and Ax1 high molecular weight glutenin subunit deficient mutant lines of elite hexaploid wheat (Triticum aestivum L.) cv. Summit

The mixing properties of the dough are critical in the production of bread and other food products derived from wheat. The high molecular weight glutenin subunits (HMW-GS) are major determinants of wheat dough processing qualities. The different alleles of the HMW-GS genes in hexaploid wheat vary in their effect on dough quality. To determine the contribution of the individual HMW-GS alleles, lines deficient in HMW-GS proteins were generated by chemical mutagenesis in the elite bread wheat Triticum aestivum cv. Summit. In this report we describe the identification and characterization of Dy10 and Ax1 deficient lines. Examination of the effect of Dy10 and Ax1 deficiency on dough rheological properties by mixography showed shorter mixing time to reach peak resistance, and weaker and less extensible doughs relative to the wild type control. This is the first time that the role of Dy10 in vivo has been examined apart from the Dx5 + Dy10 allelic pair combination.     

Waxy endosperm accompanies increased fat and saccharide contents in bread wheat (Triticum aestivum L.) grain

The contents of fat, starch, pentosan, fructan, β-glucan and several mono- and oligosaccharides in grain were evaluated to find out the possible effects of the Wx-D1 gene of bread wheat using two sets of near-isogenic waxy and non-waxy lines and two low-amylose mutant lines with a common genetic background of Kanto 107. These materials have two non-functional Wx-A1b and Wx-B1b alleles in common. Waxy near-isogenic lines with a non-functional Wx-D1d allele showed consistently increased contents of fat, total fructan, β-glucan, glucose, fructose, sucrose, 1-kestose, 6-kestose, neokestose, nystose and bifurcose compared with non-waxy lines with a functional Wx-D1a allele throughout three growing/harvest seasons. Starch and total pentosan contents were inconsistently influenced by the allelic status of the Wx-D1 locus, while water-soluble pentosan and raffinose contents were not affected. The compositional changes of a low-amylose mutant line with an almost non-functional Wx-D1f allele were closely similar to those of waxy near-isogenic lines, while significantly different changes were barely observed in another low-amylose mutant line with a partly functional Wx-D1g allele in two seasons. These results showed that the Wx-D1 gene has pleiotropic effects on the fat and saccharide contents of bread wheat grain.     

New B low Mr glutenin subunit alleles at the Glu-A3, Glu-B2 and Glu-B3 loci and their relationship with gluten strength in durum wheat

The B low M_r subunits of glutenin of the F₂ generation from three durum wheat crosses were analysed. Three new alleles were found at three different loci: Glu-A3i coding for 5+20 subunits, Glu-B2c coding for subunit 12* and Glu-B3l coding for 1+3+13*+16 subunits. The genetic distances between Glu-A3-Gli-A1, Glu-B2-Gli-B1, Glu-B3-Glu-B2 and Glu-B3-Gli-B1 were calculated. The effects of the allelic variation at the Glu-A3, Glu-B2 and Glu-B3 on protein content and gluten strength, as measured by the SDS-sedimentation test, were determined using F₄ lines from the three crosses. All the new alleles affected significantly gluten strength. The presence of Glu-A3i had a negative influence on SDSS values compared with the allele a. For Glu-B2 and Glu-B3 the data obtained enable the effects of the alleles on SDSS volume to be ranked: a=b>c for Glu-B2 and a>b>l for Glu-B3. The results also shown that the allelic variants at Glu-B3 had a much greater effect on gluten strength than the variants at Glu-A3 or Glu-B2 loci. A high percentage of variation in sedimentation volume was explained by the prolamins (52 and 70%).     

Development of introgression lines with 18 alleles of glutenin subunits and evaluation of the effects of various alleles on quality related traits in wheat (Triticum aestivum L.)

In this report, we present a set of 104 ILs with 18 alleles for five glutenin loci. They were developed from crossing and backcrossing 64 varieties as donor parents to Yanzhan 1 as recurrent parent. The effects of the 18 alleles on nine dough quality parameters were evaluated in a similar background using these lines. The results showed that Glu-A1a produced the highest SDS-sedimentation volume (Ssd), midline time x=8 width (MTxW), mixing tolerance (MT) and the lowest weakening slope (WS). At the Glu-B1 locus, Glu-B1f produced the highest values for all quality parameters but WS. At the Glu-D1 locus, Glu-D1d was the best for Ssd, grain hardness (GH), midline peak width (MPW), MTxW and MT. The positive effects of Glu-B1f on GH and Glu-B3b on Ssd were mainly from the effect of GPC. Overall, 5 interactive loci and 13 interactive alleles were found to be significant. No negative interaction between high quality glutenin alleles was detected. The preferred allele combinations for breeding were recommended based on the additive and interactive effects. Our results suggest that the ILs with multiple alleles are ideal genomic stocks for evaluating the effects of alleles on some traits and for pyramiding favorable alleles in breeding wheat varieties.     

Effects of different prolamin alleles on durum wheat quality properties

The F₄ progenies of four durum wheat crosses were used to determine the effects of different prolamin alleles on quality properties evaluated by the SDS sedimentation, mixograph, micro-alveograph and vitreousness tests and by protein content. Allelic compositions of the gliadins (Gli-B1 and Gli-2 loci) and the glutenins (Glu-1, Glu-3 and Glu-B2 loci) were determined. Alleles at the Glu-B3 locus showed a strong influence on quality measured by SDSS, mixograph and alveograph tests. Significant interactions between Glu-B3 and other glutenin loci were also detected. Prolamin composition explained more than 30% of the variation in SDSS, mixograph MT and alveograph W. The mixograph parameter BDR, and alveograph P and L parameters were the most erratic with between 8 and 76% of variation explained by prolamin composition. In general, no significant associations of prolamins with vitreousness or protein content were found. A significant correlation was detected between SDSS, MT and W. These results together with those from previous studies have important implications for wheat breeders since selection based on good alleles at Glu-B3 (a, c, j) together with favourable alleles at other loci such as Glu-A1 (subunit 1), Glu-A3 (a, c, d, h), Glu-B2 (a,b) and Gli-B1 (ω-35) could improve durum wheat quality.     

利用HPCE构建小麦LMW-GS标准图谱初探

为给不同小麦品种低分子量谷蛋白亚基（LMW-GS）的鉴定、品质预测提供快速、准确的方法,以黄淮麦区14个普通小麦品种为材料,首先利用SDS-PAGE和分子标记确定LMW-GS组成,然后采用改进的HPCE体系分离LMW-GS,最后通过控制变量法比对小麦品种间相同亚基以判断各LMW-GS亚基在图谱中对应的峰。结果表明,采用SDS-PAGE和分子标记相结合的方法可以更加准确地鉴定小麦LMW-GS;利用HPCE体系获得的小麦LMW-GS图谱具有很高的重复性,重复试验所得小麦LMW-GS图谱中各个峰迁移时间的相对标准偏差均在0.30%以下;通过控制变量法所建立的小麦LMW-GS图谱可以实现Glu-A3a、Glu-A3c、Glu-A3d、Glu-B3a、Glu-B3d、Glu-B3h的快速鉴定。