Next Article in Journal
Studies on the Yield and Chemical Composition of the Herb of Plants of the Genus Ocimum Depending on the Development Stage of the Plant
Previous Article in Journal
The Effect of Neutral Salt and Alkaline Stress with the Same Na+ Concentration on Root Growth of Soybean (Glycine max (L.) Merr.) Seedlings
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Agronomy
Volume 12
Issue 11
10.3390/agronomy12112709
agronomy-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Identification and Introgression of a Novel HMW-GS Gene from Aegilops tauschii

by
Cunyao Bo
1,†,
Zhongqing Fan
2,†,
Xin Ma
1,
Anfei Li
1,
Hongwei Wang
1,
Lingrang Kong
1 and
Xiaoqian Wang
1,*
1
State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai’an 271018, China
2
Shandong Nongda Fertilizer Sci & Tech Co., Ltd., Tai’an 271000, China
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Agronomy 2022, 12(11), 2709; https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12112709
Submission received: 6 September 2022 / Revised: 25 October 2022 / Accepted: 28 October 2022 / Published: 1 November 2022
Browse Figures
Review Reports Versions Notes

Abstract

:
High molecular weight glutenin subunits (HMW-GSs) play a major role in determining the dough quality of wheat. As the D genome donor of hexaploid wheat, Aegilops tauschii is an important genetic resource for wheat quality breeding. In the present study, a novel HMW-GSs from Ae. tauschii was identified and designated as Glu-Dt1. Multiple sequence alignment indicated that one cysteine was mutated into arginine in the y-type subunit. Site-directed mutagenesis technology was applied to verify the function of gene Glu-Dt1. Three introgression lines (ILs), B9, B25, and B35 with the Glu-D1 loci substituted by Glu-Dt1 were detected from the BC3F5 population derived from hexaploid wheat cultivar Jimai22 and Ae. tauschii Y215 through the direct hybridization approach. The dough quality and agronomic performance analysis were performed, which provide valuable resources for wheat genetic studies and breeding for distinctive end-use quality.

1. Introduction

Wheat (Triticum aestivum L.) is a major cereal contributing to the nutrition of humankind [1]. Gluten proteins, the main storage proteins in wheat, are major determinants of wheat end-use quality [2]. High molecular weight glutenin subunits (HMW-GSs) account for only 7–15% of wheat gluten proteins. However, they explain 45–70% of the variation in gluten and end-use parameters [3,4,5], which are key factors in determining gluten’s elasticity [6]. In common wheat, HMW-GSs are encoded by genes at Glu-1 loci on the long arms of the homologous group 1 chromosomes (1A, 1B, and 1D), and each locus consists of two tightly linked genes encoding an x-type subunit with a larger molecular weight and a y-type subunit with a smaller one, respectively [7,8]. Due to allelic variation and gene silencing, hexaploid wheat cultivars usually express three to five HMW-GSs, but the composition of HMW-GSs often differs among different varieties [7,9,10].
HMW-GSs is comprised of non-repetitive N- and C-terminal domains flanking a central repetitive domain, which confers the elasticity to gluten [11,12]. The repetitive domain contains 60–80% of the entire amino acid sequence and is rich in glutamine, proline, and glycine and poor in sulfur (0 or 1 cysteine) [13]. The N- and C-terminal domains are rich in charged residues and contain most of the cysteine residues [11]. The N-terminal domain contains three to five cysteine residues, while the C-terminal domain has only one. Cysteine residues provide intermolecular disulphide bonds between HMW-GS and LMW-GS to form protein polymers [14]. The number and positions of cysteine residues available for intermolecular bonds affects the size of gluten polymers and dough properties [13,15].
It is believed that the D-subgenome locus Glu-D1 has a main effect on wheat quality globally [13]. However, limited variation at the Glu-D1 locus in hexaploid wheat was found because very few accessions participated in the origin of hexaploid wheat [7,9]. Aegilops tauschii (2n = 2x = 14, DD) is the diploid D genome donor of hexaploid wheat [16]. The genes for storage proteins have higher genetic diversity in Ae. tauschii compared with hexaploid wheat, which are valuable resources for the improvement of wheat quality [17]. Several favorable genes for the storage protein have been identified in Ae. tauschii [18,19,20,21,22,23,24]. However, few of them were introgressed into wheat and applied in wheat breeding. The “direct hybridization” approach was an effective way to introgress desired D genome regions into hexaploid wheat without disrupting desirable allele combinations in the bread wheat A and B subgenomes [16,25,26]. This approach has been widely utilized to improve pathogen and pest resistance [26,27,28]. However, few studies have been reported for the utilization of the “direct hybridization” approach in the grain quality and grain yield of wheat [29].
In the present study, a novel HMW-GSs gene was cloned from Ae. tauschii and successfully transferred from Ae. tauschii into an elite winter wheat through the direct hybridization approach. This study will provide new resources for wheat genetic study and quality breeding.

2. Materials and Methods

2.1. Plant Material

Initially, the high yield winter wheat cultivar Jimai22 with HMW-GS alleles of 1, 7 + 8, and 2 + 12 on the Glu-A1, Glu-B1 and Glu-D1 loci, respectively, was crossed with Ae. tauschii Y215, and the F1 was generated by embryo rescue. Three subsequent backcrosses to Jimai22 were performed to get the BC3F1 population. The further successive self-fertilization was conducted for four generations to generate a BC3F5 population by the single seed descent method with the population size being approximately 500. Three introgression lines (ILs), B9, B25, and B35 with the Glu-D1 loci substituted by Glu-Dt1 in Y215 were detected from the BC3F5 population.

2.2. SDS-PAGE Analysis

Seed protein extraction and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were performed as described by Yan et al. [30]. Briefly, total proteins were extracted from crushed single seeds and mixed with 60% ethanol (v/v) for 30 min to remove gliadins. The extraction buffer containing 1 g DTT, 4 g SDS, 12.5 mL 1 M Tris-HCl (pH 6.8), 20 mL glycerol and 50 mL deionized water were then prepared to extract glutenins. The supernatants were used for SDS-PAGE analysis, which was performed on 10% running gel [31].

2.3. Cloning the ORFs of HMW-GSs

Genomic DNA was extracted from young leaves of Ae. tauschii Y215 using a Plant Genomic DNA kit (Tiangen, Beijing, China) according to the manufacturer’s instructions. The HMW-GS gene Glu-Dt1 was cloned from Ae. tauschii using degenerate oligonucleotide primers PF1 and PR1 (Table S1). PCR was performed in a total volume of 50 µL containing 2.5 U LA Taq with GC buffer I (TaKaRa, Dalian, China). The PCR cycle consisted of an initial 4 min denaturation at 94 °C, followed by 36 cycles at 94 °C for 50 s, at 68 °C for 4 min, and at 68 °C for 10 min. A fragment amplified by PCR was purified on 1% agarose gel and cloned into a pLUG-Prime® TA-Cloning vector (GenetBio, Korea) in Escherichia coli strain DH5α (Tiangen, Beijing, China). The cloned fragment was sequenced by the Beijing Genomics Institute. To minimize errors, at least three individual clones were sequenced.

2.4. The Dough Quality Analysis of ILs

A 10 g-Mixograph (National Mfg, Corp., Lincoln, NE, USA) was used to analyze dough rheological properties according to the AACC 54-40A method, and the following parameters were recorded: MPT, midline peak time (min); MPH, midline peak height (%); WP, width of peak time (%); MT × W: width at eight minutes (%). The grain protein content, wet gluten, development time, and stable time were analyzed by a DA7200 near-infrared spectroscope (Perten, Sweden).
The physical and textural properties of Chinese steamed bread (CSB) were determined according to Li et al. [32] and Zhang, et al. [33] with some modifications. In brief, 500 g flour, 200 g water and 4 g yeast were mixed in a mixing machine (SZM5, Xuzhong Co., Ltd., Guangzhou, China), and fermented at 30 °C with 85% relative humidity for 15 min. The dough was then remixed for 15 min and split into three pieces of 125 g. Each portion was rounded and shaped into balls by hand. The dough were then stayed for 15 min (35 °C, 85% RH) and steamed for 25 min. After cooling for 30 min at room temperature, the CSB quality was evaluated. The volume of cooled steamed bread was determined by the rapeseed displacement method; the specific volume of steamed bread was calculated by dividing loaf volume by loaf weight according to Ma et al. [34]. Hardness, springiness, cohesiveness, gumminess, chewiness and resilience of the CSB were determined using a TA-XT2 Texture Analyzer (Stable Micro Systems, Surrey, England) equipped with a 25 mm diameter aluminum cylindrical probe. The test settings were as follows: pretest speed 3.0 mm/s; test speed: 1 mm/s; posttest speed: 5 mm/s; the deformation level was 40% of the original height.

2.5. The Agronomic Performance of ILs

The recurrent parent Jimai22 and three ILs B9, B25, and B35 were grown in Heze (E1), Taian (E2), and Dezhou (E3) of Shandong Province during 2014–2015 and 2015–2016. Each line was planted in a two-row plot with 50 seeds per row with a row length of 2.0 m and a row spacing of 0.25 m. Two replicates were performed under each environment. Heading date (HD, in days) or flowering date (FD) were visually recorded for each line when ~50% of plants had headed or flowered and were represented as days from sowing to heading or flowering. At the physiological maturity stage, the middle five uniform plants in each plot were selected to evaluate plant height (PH, in cm), spike length (SL, in cm), spikelet number per spike (SPS), and grain number per spike (GN). SL was measured in centimeters from the base of the first solid spikelet to the top of the spikelet excluding awns. SPS and GN was recorded by counting the number of spikelets and grain of the main spike. Ten middle plants in the two rows were harvested to measure the thousand grain weight (TGW, in g) using a grain seed measurement machine (SC-E, Wanshen Technology Company, Hangzhou, China). The average data of two years for each environment were utilized for data analysis.

2.6. Site-Directed Mutagenesis of Glu-Dt1 Genes

The development of reduction-oxidation methods have been proved to be an effective method to investigate the effects of glutenin subunits on dough characteristics [35,36,37]. To verify the functionality of this amino acid substitution, site-directed mutagenesis was carried out using the Fast Mutagenesis System (Transgen, Beijing) according to Ma et al. [35]. In brief, the primers PF2 and PR2 were designed to clone the CDS (coding region) of Glu-Dt1 without the signal peptide coding sequence. The CDS was then cloned into a pMD18-T vector. The primers PF3 and PR3 were designed to amplify the pMD18-T vector containing Glu-Dt1 (Table S1). The PCR amplification products were treated with DMT enzyme at 37 °C for 1 h and then transformed into DH5a. The mutation sites were sequenced to confirm the altered sites.

2.7. SDS Sedimentation Test

The SDS sedimentation test was performed according to the method described by Carter et al. [38], with some minor modifications. Briefly, the base flour of wheat cv. Jimai 22 (3.0 g) was incorporated with 3.0 mg purified proteins and incubated in 50 mL of distilled water in a 100 mL measuring cylinder for 30 min at 37 °C. The cylinder was shaken for 30 s every 5 min. Then, 50 mL lactic acid-SDS solution was added, and the cylinder was shaken for 3 min twice with resting for 5 min in the middle. The cylinders were left in an upright position for 5 min, and the height (mm) of the sediment was recorded. Each test was performed twice.

2.8. Analysis of Solvent Retention Capacity (SRC)

The SRC test was employed according to the Approved AACC Method 56-11 [39] with some minor modifications. The 50 mg purified proteins were incorporated with 10 g base flour of wheat Jimai 22. Four different solvents (distilled water, 5% sodium carbonate w/w, 50% sucrose w/w, and 5% lactic acid w/w) were used to determine the four SRC values. The 1 g incorporated flour was placed into a 10 mL centrifuge tube, to which 5 mL of the appropriate solvent was added. The tubes were vortex-mixed until all of the flour was suspended. The tubes were horizontally shaken for 20 min at 1 min intervals every 5 min to allow the samples to solvate and swell. Tubes were then centrifuged at 1200× g for 10 min at room temperature. After centrifugation, the supernatant was discarded and the wet pellet was dried for 10 min and weighed. SRC values were calculated as in Haynes et al. [40]. All SRC analyses were performed in duplicate.

3. Results

3.1. Cloning the HMW-GS Genes from Ae. tauschii

The complete open reading frames (ORFs) encoding the HMW-GSs were amplified. The size of the cloned genes were 2400 bp and 1308 bp for the x-type and y-type subunits, respectively. Analysis of the predicted amino acid sequence of Glu-Dt1 indicated that four and six cysteine residues were present in the x-type subunit and y-type subunit, respectively. Multiple sequence alignments indicated that one cysteine was mutated into arginine in the y-type subunit, and no significant difference was found in the x-type subunit (Figure 1).

3.2. The Construction and Performance of ILs

SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) analysis showed that the three ILs, B9, B25, and B35 with the Glu-D1 loci substituted by Glu-Dt1 in Ae. tauschii Y215 were detected from the BC3F5 population (Figure 2a). The x-type and y-type subunits migrated slower than that of Jimai22, respectively (Figure 2a).
To detect the influence on dough quality of the substitution in Glu-D1 loci, several tests were performed for three ILs and Jimai22. Compared with Jimai22, three ILs had significantly lower MPT (midline peak time) and MT × W (width at eight minutes), higher grain protein content, and lower stable time (Table 1; Figure 3). These three ILs showed significantly higher loaf height, volume and specific volume than Jimai22. Texture analysis showed that springiness and cohesiveness were significantly increased and the hardness, gumminess, chewiness and resilience were significantly decreased in the three ILs than in Jimai22 (Table 2).
Compared with Jimai22, B9 showed significantly higher PH and lower SL, SPS, GN, and TKW. B25 showed significantly higher HD, FD, PH, and TKW and significantly lower GN than Jimai22. B35 have significantly higher HD, FD, PH, SL, SPS, GN, and TGW than Jimai22 (Table 3, Figure 2b).

3.3. Site-Directed Mutagenesis of Glu-Dt1 Coding Genes

The amino acid analysis showed that one cysteine was mutated into arginine in y-type subunit of Glu-Dt1 (Figure 1). The site-directed mutagenesis was carried out to verify the functionality of this amino acid substitution. The SRC values increased more for the incorporated flour of Mut Y215-Y than the incorporated flour of Y215-Y (Figure 4a). The SDS sedimentation volume for incorporated flour of Y215-Y and Mut Y215-Y were 16.51 mL and 25.10 mL, respectively, which showed that Mut Y215-Y was more effective in promoting SDS sedimentation volume than Y215-Y (Figure 4b). This result indicated that the absence of cysteine residue in Y215-Y had significant impact on the properties of the dough.

4. Discussion

Compared with the diploid ancestor Ae. tauschii, hexaploid wheat has a broader adaptation and extended potential to make diverse food products with the addition of D-subgenome. The D-subgenome was regarded as the major driver of bread quality [41]. However, the polyploidization with few Ae. tauschii accessions that participated in the origin of the hexaploid species imposed a genetic bottleneck, which resulted in the limited genetic diversity in the wheat D genome [9,42]. Ae. tauschii is a reservoir for quality improvement in wheat breeding [9,12]. Previous studies have shown that HMW-GSs from the Ae. tauschii have a significant influence on dough properties in synthetic hexaploid wheats [9,43]. Thus, exploiting the favorable alleles of Ae. tauschii could benefit the revelation of the mechanism of wheat storage proteins and accelerate the breeding process of wheat.
As an important source of carbohydrates worldwide, wheat is the most complex cereal in terms of end-use quality. Wheat flour can be made into limitless categories of foods including breads, steamed breads, crackers, cookies, cakes, noodles, pasta, and couscous [44]. The evaluation criterion to judge the quality of a particular flour should depend on consumer preferences, the product and its processing [45]. For example, the flour with high dough quality generally has high bread baking quality, while also exhibiting poor cookie- or cake-baking quality [46]. The flour with medium dough quality generally has high steam bread quality. Therefore, the wheat variety with diverse dough quality should be cultivated to meet the distinctive end-use quality. In this study, a novel gene for HMW-GSs, Glu-Dt1, was cloned from Ae. tauschii, and was successfully transferred into elite wheat cultivar Jimai22 through the direct hybridization approach. The ILs showed lower dough quality but improved CSB quality. In particular, B35 showed significantly higher SL, SPS, GN, and TGW than Jimai22, which can serve as a precious resource for further breeding and genetic studies in wheat.
The cysteine contents of HMW subunits could affect the formation of intermolecular disulfide bonds and hence properties of the glutenin polymers [47]. Previous studies have demonstrated that x-type subunits and y-type subunits usually contain four and seven conserved cysteine residues, respectively [14,35]. Site-directed mutagenesis technology has been widely used for verifying the function of genes [35,37]. In the present study, the substitution from cysteine to arginine in the y-type subunit could be responsible for the lower dough quality by decreasing the number and affecting the pattern of disulphide cross-links in the gluten in polymers, which have been verified by site-directed mutagenesis technology. Previous studies have suggested that the number and distribution of cysteine residues were two of the key factors influencing wheat dough quality [18,34,48,49]. Although exploiting the new HMW-GSs genes from wild species was an effective way to broaden the genetic diversity of wheat [8,21,31,35,50,51], the gene engineering could be utilized to improve the dough quality by modifying the number or the structure of cysteine residues in HMW subunits [11,35].

5. Conclusions

The HMW-GSs in bread wheat are major determinants of the viscoelastic properties of dough. To enrich the genetic diversity of HMW-GSs in wheat, a novel HMW-GSs designated as Glu-Dt1 from Ae. tauschii was identified. Multiple sequence alignment indicated that one cysteine was mutated into arginine in the y-type subunit, and no significant difference was found in the x-type subunit. Site-directed mutagenesis was conducted to investigate the influence of this cysteine residue substitution on wheat dough quality. By the direct hybridization approach, three ILs, B9, B25, and B35, with the Glu-D1 loci substituted by Glu-Dt1, were developed. The dough quality and agronomic performance analysis were performed, which provide valuable information for wheat breeding.

Supplementary Materials

The following supporting information can be downloaded at: https://0-www-mdpi-com.brum.beds.ac.uk/article/10.3390/agronomy12112709/s1, Table S1: The primers used in this study.

Author Contributions

Conceptualization, H.W. and L.K.; Methodology, X.M.; Software, X.W.; Validation, C.B., Z.F. and A.L.; Formal Analysis, X.W.; Investigation, C.B. and Z.F.; Resources, H.W. and L.K.; Data Curation, Z.F.; Writing—Original Draft Preparation, X.W. and C.B.; Writing—Review & Editing, H.W. and X.W.; Visualization, C.B.; Supervision, L.K.; Project Administration, X.W.; Funding Acquisition, X.W. All authors have read and agreed to the published version of the manuscript.

Funding

This study was funded by the National Natural Science Foundation of China (31901491).

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Conflicts of Interest

The authors declare that they have no conflict of interest.

References

  1. Kiszonas, A.M.; Morris, C.F. Wheat breeding for quality: A historical review. Cereal Chem. 2018, 95, 17–34. [Google Scholar] [CrossRef] [Green Version]
  2. Goesaert, H.; Brijs, K.; Veraverbeke, W.S.; Courtin, C.M.; Gebruers, K.; Delcour, J.A. Wheat flour constituents: How they impact bread quality, and how to impact their functionality. Trends. Food Sci. Technol. 2005, 16, 12–30. [Google Scholar] [CrossRef]
  3. He, Z.H.; Liu, L.; Xia, X.C.; Liu, J.J.; Peña, R.J. Composition of HMW and LMW glutenin subunits and their effects on dough properties, pan bread, and noodle quality of Chinese bread wheats. Cereal Chem. 2005, 82, 345–350. [Google Scholar] [CrossRef]
  4. Li, Z.; Deng, C.; Li, H.; Liu, C.; Bian, K. Characteristics of remixed fermentation dough and its influence on the quality of steamed bread. Food Chem. 2015, 179, 257–262. [Google Scholar] [CrossRef] [PubMed]
  5. Wang, D.; Zhang, K.; Dong, L.; Dong, Z.; Li, Y.; Hussain, A.; Zhai, H. Molecular genetic and genomic analysis of wheat milling and end-use traits in China:Progress and perspectives. Crop J. 2018, 6, 68–81. [Google Scholar] [CrossRef]
  6. Cornish, G.B.; Békés, F. Prediction of Dough Properties for Bread Wheats. In Gliadin and Glutenin: The Unique Balance of Wheat Quality; Cereals & Grains Association: Saint Paul, MN, USA, 2006; pp. 243–280. [Google Scholar]
  7. Payne, P.I.; Lawrence, G.J. 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. 1983, 11, 29–35. [Google Scholar]
  8. Yan, M.; Li, M.; Yang, Z.; Yu, F.; Du, X. Characterization of 1Sty13, a novel high-molecular-weight glutenin subunit from Elymus sibiricus L. Plant Genet. Resour. 2021, 19, 308–311. [Google Scholar] [CrossRef]
  9. Delorean, E.; Gao, L.L.; Lopez, J.; Consortium, T.; Poland, J. High molecular weight glutenin gene diversity in Aegilops tauschii demonstrates unique origin of superior wheat quality. Commun. Biol. 2021, 4, 1242. [Google Scholar] [CrossRef]
  10. Payne, P.I.; Law, C.N.; Mudd, E.E. Control by homoeologous group 1 chromosomes of the high-molecular-weight subunits of glutenin, a major protein of wheat endosperm. Theor. Appl. Genet. 1980, 58, 113–120. [Google Scholar] [CrossRef]
  11. Anjum, F.M.; Khan, M.R.; Din, A.; Saeed, M.; Pasha, I.; Arshad, M.U. Wheat gluten: High molecular weight glutenin subunits—Structure, genetics, and relation to dough elasticity. J. Food Sci. 2007, 72, R56–R63. [Google Scholar] [CrossRef]
  12. Gianibelli, M.C.; Larroque, O.R.; MacRitchie, F.; Wrigley, C.W. Biochemical, genetic, and molecular characterization of wheat glutenin and its component subunits. Cereal Chem. 2001, 78, 635–646. [Google Scholar] [CrossRef]
  13. Wang, J.J.; Liu, G.; Huang, Y.B.; Zeng, Q.H.; Hou, Y.; Li, L.; Ou, S.; Zhang, M.; Hu, S.Q. Dissecting the disulfide linkage of the N-Terminal domain of HMW 1Dx5 and its contributions to dough functionality. J. Agric. Food Chem. 2017, 65, 6264–6273. [Google Scholar] [CrossRef] [PubMed]
  14. Shewry, P.R.; Halford, N.G.; Tatham, A.S. High molecular weight subunits of wheat glutenin. J. Cereal Sci. 1992, 15, 105–120. [Google Scholar] [CrossRef]
  15. Pirozi, M.R.; Margiotta, B.; Lafiandra, D.; Macritchie, F. Composition of polymeric proteins and bread-making quality of wheat lines with allelic HMW-GS differing in number of cysteines. J. Cereal Sci. 2008, 48, 117–122. [Google Scholar] [CrossRef]
  16. Gaurav, K.; Arora, S.; Silva, P.; Sánchez-Martín, J.; Horsnell, R.; Gao, L.; Brar, G.S.; Widrig, V.; John Raupp, W.; Singh, N.; et al. Population genomic analysis of Aegilops tauschii identifies targets for bread wheat improvement. Nat. Biotechnol. 2021, 3, 422–431. [Google Scholar] [CrossRef]
  17. Lafiandra, D.; Masci, S.; D’Ovidio, R.; Tanzarella, O.A.; Porceddu, E.; Margiotta, B. Relationship between the D genome of hexaploid wheats (AABBDD) and Ae. squarrosa as deduced by seed storage proteins and molecular marker analyses. Hereditas 1992, 116, 233–238. [Google Scholar] [CrossRef]
  18. An, X.L.; Li, X.H.; Xiong, X.J.; Yan, Y.M.; Zhang, Y.Z.; Gao, L.Y.; Wang, A.L.; Wang, K.; Zeller, F.J.; Hsam, S.L.K. Identification and isolation of a new x-type HMW glutenin subunit 1Dx1.6 t gene from Aegilops tauschii. Plant Breed. 2009, 128, 41–45. [Google Scholar] [CrossRef]
  19. Pflüger, L.A.; D’Ovidio, R.; Margiotta, B.; Peña, R.; Mujeeb-Kazi, A.; Lafiandra, D. Characterisation of high- and low-molecular weight glutenin subunits associated to the D genome of Aegilops tauschii in a collection of synthetic hexaploid wheats. Theor. Appl. Genet. 2001, 103, 1293–1301. [Google Scholar] [CrossRef]
  20. Rasheed, A.; Mahmood, T.; Kazi, A.G.; Ghafoor, A.; Mujeeb-Kazi, A. Allelic variation and composition of HMW-GS in advanced lines derived from d-genome synthetic hexaploid / bread wheat (Triticum aestivum L.). J. Crop Sci. Biotechnol. 2012, 15, 1–7. [Google Scholar] [CrossRef]
  21. Wang, K.; An, X.L.; Pan, L.P.; Dong, K.; Gao, L.Y.; Wang, S.L.; Xie, Z.Z.; Zhang, Z.; Appels, R.; Ma, W.; et al. Molecular characterization of HMW-GS 1Dx3t and 1Dx4t genes from Aegilops tauschii and their potential value for wheat quality improvement. Hereditas 2012, 149, 41–49. [Google Scholar] [CrossRef]
  22. Yan, Y.M.; Hsam, S.L.K.; Yu, J.Z.; Jiang, Y.; Zeller, F.J. Genetic polymorphisms at Gli-Dt gliadin loci in Aegilops tauschii as revealed by acid polyacrylamide gel and capillary electrophoresis. Plant Breed. 2003, 122, 120–124. [Google Scholar] [CrossRef]
  23. Yan, Y.; Hsam, S.L.K.; Yu, J.; Jiang, Y.; Zeller, F.J. Allelic variation of the HMW glutenin subunits in Aegilops tauschii accessions detected by sodium dodecyl sulphate (SDS-PAGE), acid polyacrylamide gel (A-PAGE) and capillary electrophoresis. Euphytica 2003, 130, 377–385. [Google Scholar] [CrossRef]
  24. Zhang, Y.; Li, X.; Wang, A.; An, X.; Zhang, Q.; Pei, Y.; Gao, L.; Ma, W.; Appels, R.; Yan, Y. Novel x-type high-molecular-weight glutenin genes from Aegilops tauschii and their implications on the wheat origin and evolution mechanism of Glu-D1-1 proteins. Genetics 2008, 178, 23–33. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  25. Cox, T.S.; Wu, J.; Wang, S.; Cai, J.; Zhong, Q.; Fu, B. Comparing two approaches for introgression of germplasm from Aegilops tauschii into common wheat. Crop J. 2017, 5, 355–362. [Google Scholar] [CrossRef]
  26. Zhang, D.; Zhou, Y.; Zhao, X.; Lv, L.; Zhang, C.; Li, J.; Sun, G.; Li, S.; Song, C.-P. Development, utilization of introgression lines using synthetic octaploid wheat (Aegilops tauschii× hexaploid wheat) as donor. Front. Plant Sci. 2018, 9, 1113. [Google Scholar] [CrossRef]
  27. Olson, E.L.; Rouse, M.N.; Pumphrey, M.O.; Bowden, R.L.; Gill, B.S.; Poland, J.A. Introgression of stem rust resistance genes SrTA10187 and SrTA10171 from Aegilops tauschii to wheat. Theor. Appl. Genet. 2013, 126, 2477–2484. [Google Scholar] [CrossRef]
  28. Raupp, W.J.; Sukhwinder-Singh; Brown-Guedira, G.L.; Gill, B.S. Cytogenetic and molecular mapping of the leaf rust resistance gene Lr39 in wheat. Theor. Appl. Genet. 2001, 102, 347–352. [Google Scholar] [CrossRef]
  29. Cox, T.S.; Sears, R.G.; Bequette, R.K. Use of winter wheat x Triticum tauschii backcross populations for germplasm evaluation. Theor. Appl. Genet. 1995, 90, 571–577. [Google Scholar] [CrossRef]
  30. Yan, Z.; Wan, Y.; Liu, K.; Zheng, Y.; Wang, D. Identification of a novel HMW glutenin subunit and comparison of its amino acid sequence with those of homologous subunits. Chin. Sci. Bull. 2002, 47, 222–226. [Google Scholar] [CrossRef]
  31. Ma, C.; Yang, Y.; Li, X.; Ge, P.; Guo, G.; Subburaj, S.; Zeller, F.J.; Hsam, S.L.K.; Yan, Y. Molecular cloning and characterization of six novel HMW-GS genes from Aegilops speltoides and Aegilops kotschyi. Plant Breed. 2013, 132, 284–289. [Google Scholar] [CrossRef]
  32. Li, Y.; An, X.; Yang, R.; Guo, X.; Yue, G.; Fan, R.; Li, B.; Li, Z.; Zhang, K.; Dong, Z.; et al. Dissecting and enhancing the contributions of high-molecular-weight glutenin subunits to dough functionality and bread quality. Mol. Plant 2015, 8, 332–334. [Google Scholar] [CrossRef] [PubMed]
  33. Zhang, P.; Jondiko, T.O.; Tilley, M.; Awika, J.M. Effect of high molecular weight glutenin subunit composition in common wheat on dough properties and steamed bread quality. J. Sci. Food Agric. 2014, 94, 2801–2806. [Google Scholar] [CrossRef] [PubMed]
  34. Ma, S.; Li, L.; Wang, X.; Zheng, X.; Bian, K.; Bao, Q. Effect of mechanically damaged starch from wheat flour on the quality of frozen dough and steamed bread. Food Chem. 2016, 202, 120–124. [Google Scholar] [CrossRef] [PubMed]
  35. Ma, X.; Du, X.; Bo, C.; Wang, H.; Li, A.; Kong, L. Modification of a novel x-type high-molecular-weight glutenin subunit gene from Aegilops markgrafii to improve dough strength of wheat flour. Crop Pasture Sci. 2018, 69, 873–878. [Google Scholar] [CrossRef]
  36. Anderson, O.D.; Bekes, F.; D’Ovidio, R. Effects of specific domains of high-molecular-weight glutenin subunits on dough properties by an in vitro assay. J. Cereal Sci. 2011, 54, 280–287. [Google Scholar] [CrossRef]
  37. Gao, X.; Zhang, Q.; Newberry, M.P.; Chalmers, K.J.; Mather, D.E. A cysteine in the repetitive domain of a high-molecular-weight glutenin subunit interferes with the mixing properties of wheat dough. Amino Acids 2013, 44, 1061–1071. [Google Scholar] [CrossRef]
  38. Carter, B.P.; Morris, C.F.; Anderson, J.A. Optimizing the SDS sedimentation test for end-use quality selection in a soft white and club wheat breeding program. Cereal Chem. 1999, 76, 907–911. [Google Scholar] [CrossRef]
  39. Bettge, A.D.; Morris, C.F.; Demacon, V.L.; Kidwell, K.K. Adaptation of AACC method 56-11, solvent retention capacity, for use as an early generation selection tool for cultivar development. Cereal Chem. 2002, 79, 670–674. [Google Scholar] [CrossRef]
  40. Haynes, L.; Bettge, A.; Slade, L.; Levine, H.; Communicator, H.N.A. Soft Wheat and flour products methods review: Solvent retention capacity equation correction. Cereal Foods World 2009, 54, 174–175. [Google Scholar] [CrossRef]
  41. Dubcovsky, J.; Dvorak, J. Genome plasticity a key factor in the success of polyploid wheat under domestication. Science 2007, 316, 1862. [Google Scholar] [CrossRef] [Green Version]
  42. Wang, J.; Luo, M.C.; Chen, Z.; You, F.M.; Wei, Y.; Zheng, Y.; Dvorak, J. Aegilops tauschii single nucleotide polymorphisms shed light on the origins of wheat D-genome genetic diversity and pinpoint the geographic origin of hexaploid wheat. New Phytol. 2013, 198, 925–937. [Google Scholar] [CrossRef] [PubMed]
  43. Hsam, S.L.K.; Kieffer, R.; Zeller, F.J. Significance of Aegilops tauschii glutenin genes on breadmaking properties of wheat. Cereal Chem. 2001, 78, 521–525. [Google Scholar] [CrossRef]
  44. Wrigley, C. Chapter 5–Wheat: Grain-quality characteristics and management of quality requirements. In Cereal Grains; Woodhead Publishing: Sawston, UK, 2017; pp. 91–134. [Google Scholar]
  45. Huertas-García, A.B.; Castellano, L.; Guzmán, C.; Alvarez, J.B. Potential use of wild einkorn wheat for wheat grain quality improvement: Evaluation and characterization of Glu-1, Wx and Ha loci. Agronomy 2021, 11, 816. [Google Scholar] [CrossRef]
  46. Morris, C.F. Cereals: Overview of Uses: Accent on Wheat Grain. In Encyclopedia of Food Grains, 2nd ed.; Wrigley, C., Corke, H., Seetharaman, K., Faubion, J., Eds.; Academic Press: Oxford, UK, 2016; pp. 1–7. [Google Scholar]
  47. Shewry, P.; Gilbert, S.; Savage, A.; Tatham, A.; Wan, Y.F.; Belton, P.; Wellner, N.; D’Ovidio, R.; Békés, F.; Halford, N. Sequence and properties of HMW subunit 1Bx20 from pasta wheat (Triticum durum) which is associated with poor end use properties. Theor. Appl. Genet. 2003, 106, 744–750. [Google Scholar] [CrossRef] [PubMed]
  48. Chen, F.; Liu, S.; Zhao, F.; Xu, C.; Xia, G. Molecular characterisation of the low-molecular weight glutenin subunit genes of tall wheatgrass and functional properties of one clone Ee34. Amino Acids 2010, 38, 991–999. [Google Scholar] [CrossRef]
  49. Chen, F.; Zhao, F.; Liu, R.; Xia, G. Functional properties of two low-molecular-weight glutenin subunits carrying additional cysteine residues from hybrid introgression line II-12 derived from Triticum aestivum and Agropyron elongatum. Food Chem. 2011, 127, 1773–1776. [Google Scholar] [CrossRef]
  50. Chen, W.J.; Xing, F.; Bo, Z.; Liu, B.L.; Yan, Z.H.; Zhang, L.Q.; Yuan, Z.W.; Zheng, Y.L.; Zhang, H.G.; Liu, D.C. Novel and ancient HMW glutenin genes from Aegilops tauschii and their phylogenetic positions. Genet. Resour. Crop Evol. 2012, 59, 1649–1657. [Google Scholar] [CrossRef]
  51. Du, X.; Zhao, J.; Ma, X.I.N.; Yin, H.; Wang, H.; Li, A.; Kong, L. Cloning and characterization of novel fast ω-gliadin genes in Triticum monococcum. J. Genet. 2015, 94, 323–327. [Google Scholar] [CrossRef]
Agronomy 12 02709 g001 550
Figure 1. The alignment of amino acid sequences of HMW-GS genes for (a) x-type subunit and (b) y-type subunit. The cysteine residue is indicated by a red triangle. The mutation site is indicated by a red arrow.
Figure 1. The alignment of amino acid sequences of HMW-GS genes for (a) x-type subunit and (b) y-type subunit. The cysteine residue is indicated by a red triangle. The mutation site is indicated by a red arrow.
Agronomy 12 02709 g001
Agronomy 12 02709 g002 550
Figure 2. (a) The composition of glutenins in B9, B25, B35 and their parents. (b) Plant phenotype of B9, B25, B35 and Jimai22.
Figure 2. (a) The composition of glutenins in B9, B25, B35 and their parents. (b) Plant phenotype of B9, B25, B35 and Jimai22.
Agronomy 12 02709 g002
Agronomy 12 02709 g003 550
Figure 3. The dough rheological properties for three ILs and Jimai22. *, **, and, *** represent the significance of ANOVA at p < 0.1, p < 0.01 and p < 0.001, respectively. MPT, midline peak time (min); MPH, midline peak height (%); RPS, right of peak slope (°); WP, width of peak time (%); and MT × W, width at eight minutes (%).
Figure 3. The dough rheological properties for three ILs and Jimai22. *, **, and, *** represent the significance of ANOVA at p < 0.1, p < 0.01 and p < 0.001, respectively. MPT, midline peak time (min); MPH, midline peak height (%); RPS, right of peak slope (°); WP, width of peak time (%); and MT × W, width at eight minutes (%).
Agronomy 12 02709 g003
Agronomy 12 02709 g004 550
Figure 4. The dough properties of incorporated flour with the incorporation of purified protein Y215-Y and Mut Y215-Y. (a) SRC values; (b) SDS sedimentation volume. The * represented the significance of ANOVA at p < 0.1.
Figure 4. The dough properties of incorporated flour with the incorporation of purified protein Y215-Y and Mut Y215-Y. (a) SRC values; (b) SDS sedimentation volume. The * represented the significance of ANOVA at p < 0.1.
Agronomy 12 02709 g004
Table
Table 1. Grain protein content, wet gluten, development time, and stable time of three ILs and Jimai22 in Heze (E1), Taian (E2), and Dezhou (E3) of Shandong Province.
Table 1. Grain protein content, wet gluten, development time, and stable time of three ILs and Jimai22 in Heze (E1), Taian (E2), and Dezhou (E3) of Shandong Province.
Jimai22B9B25B35
E1E2E3E1E2E3E1E2E3E1E2E3
Grain protein content (%)13.5312.5313.5915.47 **15.46 **15.68 **13.6113.0414.1913.85 *12.94 *14.37 *
Wet gluten (%)38.237.539.844.2 **36.146.9 **39.236.740.537.038.541.9
Development time (min)2.92.83.12.5 *2.6 *2.5 *2.4 *2.92.6 *2.5 *2.4 *2.9 *
Mixograph stable time (min)2.72.73.42.2 *2.5 *2.3 **1.92.62.72.02.52.4
The * and ** suggested significance of ANOVA at p < 0.1 and p < 0.01, respectively.
Table
Table 2. Physical and textural properties of steamed bread for three introgression lines and Jimai22.
Table 2. Physical and textural properties of steamed bread for three introgression lines and Jimai22.
Parameters Jimai22B9B25B35
Loaf height (mm)45.37 ± 0.21 a53.15 ± 0.28 b50.29 ± 0.12 b51.04 ± 0.38 b
Volume (mL)126 ± 1.03 a198 ± 1.25 b186 ± 1.23 b188 ± 0.25 b
Specific volume (mL/g)1.79 ± 0.03 a2.44 ± 0.01 b2.29 ± 0.02 b2.04 ± 0.03 b
Moisture (g water/100 g sample)44.73 ± 0.01 a40.17 ± 0.01 b41.43 ± 0.02 b40.05 ± 0.03 b
Hardness (g)526.53 ± 41.13 a328.64 ± 21.92 b426.33 ± 21.13 b408.34 ± 20.73 b
Springiness (mm)0.75 ± 0.01 a0.82 ± 0.03 b0.81 ± 0.02 b0.89 ± 0.04 b
Cohesiveness (ratio)0.58 ± 0.01 a0.84 ± 0.01 b0.87 ± 0.02 b0.82 ± 0.01 b
Gumminess (g)347.09 ± 8.42 a241.45 ± 11.07 b267.18 ± 11.62 b281.43 ± 12.37 b
Chewiness (g)329.72 ± 35.34 a189.47 ± 16.32 b185.77 ± 32.04 b200.57 ± 15.33 b
Resilience (ratio)0.68 ± 0.01 a0.51 ± 0.02 b0.55 ± 0.01 b0.57 ± 0.01 b
The value in the table was the means ± standard deviations (n = 3) for each character. The letters in the same row showed the difference of ANOVA at the significant level of p < 0.05.
Table
Table 3. Agronomic performance of three ILs and Jimai22 in three environments.
Table 3. Agronomic performance of three ILs and Jimai22 in three environments.
TraitsJimai22B9B25B35
E1E2E3E1E2E3E1E2E3E1E2E3
HD200207211200206210208 **215 **219 **211 ***218 ***221 ***
FD207213219208214215215 **220 **225 **216 **224 **227 **
PH (cm)75727096 **95 **90 **80 **78 **75 **78 *76 *77 *
SL (cm)9.010.09.07.9 **8.4 **8.2 **10.110.09.110.2 ***10.5 ***10.3 ***
SPS28323016 ***20 ***18 ***30322832 **35 **34 **
GN63586150 *47 *52 *52 **49 **50 **69 **65 **67 **
TGW (g)45.0045.0245.0546.53 *46.74 *46.61 *50.65 ***49.98 ***50.47 ***48.36 *48.75 *47.96 *
(1) The *, **, and *** suggested significance of ANOVA between the corresponding ILs and Jimai22 at p < 0.1, p < 0.01, and p < 0.001, respectively. (2) E1, E2, and E3 represented Heze, Taian and Dezhou of Shandong Province, respectively. HD: heading date, FD: flowering date, PH: plant height, SL: spike length, SPS: spikelet number per spike, GN: grain number per spike, TGW: thousand grain weight.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Bo, C.; Fan, Z.; Ma, X.; Li, A.; Wang, H.; Kong, L.; Wang, X. Identification and Introgression of a Novel HMW-GS Gene from Aegilops tauschii. Agronomy 2022, 12, 2709. https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12112709

AMA Style

Bo C, Fan Z, Ma X, Li A, Wang H, Kong L, Wang X. Identification and Introgression of a Novel HMW-GS Gene from Aegilops tauschii. Agronomy. 2022; 12(11):2709. https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12112709

Chicago/Turabian Style

Bo, Cunyao, Zhongqing Fan, Xin Ma, Anfei Li, Hongwei Wang, Lingrang Kong, and Xiaoqian Wang. 2022. "Identification and Introgression of a Novel HMW-GS Gene from Aegilops tauschii" Agronomy 12, no. 11: 2709. https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12112709

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop