Abstract
Key message
Eleven wheat lines that are missing genes for the 1D-encoded omega-5 gliadins will facilitate breeding efforts to reduce the immunogenic potential of wheat flour for patients susceptible to wheat allergy.
Abstract
Efforts to reduce the levels of allergens in wheat flour that cause wheat-dependent exercise-induced anaphylaxis are complicated by the presence of genes encoding omega-5 gliadins on both chromosomes 1B and 1D of hexaploid wheat. In this study, we screened 665 wheat germplasm samples using gene specific DNA markers for omega-5 gliadins encoded by the genes on 1D chromosome that were obtained from the reference wheat Chinese Spring. Eleven wheat lines missing the PCR product corresponding to 1D omega-5 gliadin gene sequences were identified. Two of the lines contained the 1BL·1RS translocation. Relative quantification of gene copy numbers by qPCR revealed that copy numbers of 1D omega-5 gliadins in the other nine lines were comparable to those in 1D null lines of Chinese Spring, while copy numbers of 1B omega-5 gliadins were like those of Chinese Spring. 2-D immunoblot analysis of total flour proteins from the selected lines using a specific monoclonal antibody against the N-terminal sequence of omega-5 gliadin showed no reactivity in regions of the blots containing previously identified 1D omega-5 gliadins. Interestingly, RP-UPLC analysis of the gliadin fractions of the selected lines indicated that the expression of omega-1,2 gliadins was also significantly reduced in seven of the lines, implying that 1D omega-5 gliadin and 1D omega-1,2 gliadin genes are tightly linked on the Gli-D1 loci of chromosome 1D. Wheat lines missing the omega-5 gliadins encoded by the genes on 1D chromosome should be useful in future breeding efforts to reduce the immunogenic potential of wheat flour.
Similar content being viewed by others
Data availability
All reference data that are not presented in this manuscript are available in the supplementary materials.
References
Altenbach SB, Allen PV (2011) Transformation of the US bread wheat “Butte 86” and silencing of omega-5 gliadin genes. GM Crops 2:66–73. https://doi.org/10.4161/gmcr.2.1.15884
Altenbach SB, Tanaka CK, Hurkman WJ, Whitehand LC, Vensel WH, Dupont FM (2011) Differential effects of a post-anthesis fertilizer regimen on the wheat four proteome determined by quantitative 2-DE. Proteome Sci 9:46. https://doi.org/10.1186/1477-5956-9-46
Altenbach SB, Tanaka CK, Seabourn BW (2014) Silencing of omega-5 gliadins in transgenic wheat eliminates a major source of environmental variability and improves dough mixing properties of flour. BMC Plant Biol 14:393. https://doi.org/10.1186/s12870-014-0393-1
Altenbach SB, Tanaka CK, Pineau F, Lupi R, Drouet M, Beaudouin E, Morisset M, Denery-Papini S (2015) Assessment of the allergenic potential of transgenic wheat (Triticum aestivum) with reduced levels of ω5-gliadins, the major sensitizing allergen in wheat-dependent exercise-induced anaphylaxis. Agric Food Chem 63:9323–9332. https://doi.org/10.1021/acs.jafc.5b03557
Altenbach SB, Chang HC, Simon-Buss A, Jang YR, Denery-Papini S, Pineau F, Gu YQ, Huo N, Lim SH, Kang CS, Lee JY (2018) Towards reducing the immunogenic potential of wheat flour: omega gliadins encoded by the D genome of hexaploid wheat may also harbor epitopes for the serious food allergy WDEIA. BMC Plant Biol 18:291. https://doi.org/10.1186/s12870-018-1506-z
Altenbach SB, Chang HC, Yu XB, Seabourn BW, Green PH, Alaedini A (2019) Elimination of omega-1,2 gliadins from bread wheat (Triticum aestivum) flour: effects on immunogenic potential and end-use quality. Front Plant Sci 10:58. https://doi.org/10.3389/fpls.2019.00580
Altenbach SB, Chang HC, Simon-Buss A, Mohr T, Huo N, Gu YQ (2020) Exploiting the reference genome sequence of hexaploid wheat: a proteomic study of flour proteins from the cultivar Chinese Spring. Funct Integr Genomics 20:1–16. https://doi.org/10.1007/s10142-019-00694-z
Anderson OD, Gu YQ, Kong X, Lazo GR, Wu J (2009) The wheat omega-gliadin genes: structure and EST analysis. Funct Integr Genomics 9:397–410. https://doi.org/10.1007/s10142-009-0122-2
Barro F, Iehisa JC, Giménez MJ, García-Molina MD, Ozuna CV, Comino I, Sousa C, Gil-Humanes J (2016) Targeting of prolamins by RNAi in bread wheat: effectiveness of seven silencing-fragment combinations for obtaining lines devoid of coeliac disease epitopes from highly immunogenic gliadins. Plant Biotechnol J 14:986–996. https://doi.org/10.1111/pbi.12455
Bartoš P, Bareš I (1971) Leaf and stem rust resistance of hexaploid wheat cultivars salzmünder bartweizen and weique. Euphytica 20:435–440. https://doi.org/10.1007/BF00035671
Blechl A, Beecher B, Vensel W, Tanaka C, Altenbach S (2016) RNA interference targeting rye secalins alters flour protein composition in a wheat variety carrying a 1BL.1RS translocation. J Cereal Sci 68:172–180. https://doi.org/10.1016/j.jcs.2016.01.009
Camerlengo F, Sestili F, Silvestri M, Colaprico G, Margiotta B, Ruggeri R, Lupi R, Masci S, Lafiandra D (2017) Production and molecular characterization of bread wheat lines with reduced amount of α-type gliadins. BMC Plant Biol 17:248. https://doi.org/10.1186/s12870-017-1211-3
DuPont FM, Vensel WH, Encarnacao T, Chan R, Kasarda D (2004) Similarities of omega gliadins from Triticum urartu to those encoded on chromosome 1A of hexaploid wheat and evidence for their post-translational processing. Theor Appl Genet 108:1299–1308. https://doi.org/10.1007/s00122-003-1565-9
Dupont FM, Vensel WH, Tanaka CK, Hurkman WJ, Altenbach SB (2011) Deciphering the complexities of the wheat flour proteome using quantitative two-dimensional electrophoresis, three proteases and tandem mass spectrometry. Proteome Sci 9:10. https://doi.org/10.1186/1477-5956-9-10
Gil-Humanes J, Pistón F, Altamirano-Fortoul R, Real A, Comino I, Sousa C, Rosell CM, Barro F (2014) Reduced-gliadin wheat bread: an alternative to the gluten-free diet for consumers suffering gluten-related pathologies. PLoS ONE 9:e90898. https://doi.org/10.1371/journal.pone.0090898
Gil-Humanes J, Pistón F, Barro F, Rosell CM (2014) The shutdown of celiac disease-related gliadin epitopes in bread wheat by RNAi provides flours with increased stability and better tolerance to over-mixing. PLoS ONE 9:e91931. https://doi.org/10.1371/journal.pone.0091931
Huang L, Brooks SA, Li W, Fellers JP, Trick HN, Gill BS (2003) Map-based cloning of leaf rust resistance gene Lr21 from the large and polyploid genome of bread wheat. Genetics 164:655–664. https://doi.org/10.1093/genetics/164.2.655
Huo N, Zhang S, Zhu T, Dong L, Mohr T, Hu T, Liu Z, Dvorak J, Luo MC, Wang D, Lee J-Y, Altenbach S, Gu YQ (2018) Gene duplication and evolution dynamics in the homeologous regions harboring multiple prolamin and resistance gene families in hexaploid wheat. Front Plant Sci 9:673. https://doi.org/10.3389/fpls.2018.00673
Jang YR, Cho K, Kim S, Sim JR, Lee SB, Kim BG, Gu YQ, Altenbach SB, Lim SH, Goo TW, Lee JY (2020) Comparison of MALDI-TOF-MS and RP-HPLC as rapid screening methods for wheat lines with altered gliadin compositions. Front Plant Sci 11:600489. https://doi.org/10.3389/fpls.2020.600489
Jang YR, Kim S, Sim JR, Lee SB, Lim SH, Kang CS, Choi C, Goo TW, Lee JY (2021) High-throughput analysis of high-molecular weight glutenin subunits in 665 wheat genotypes using an optimized MALDI-TOF-MS method. 3 Biotech 11:92. https://doi.org/10.1007/s13205-020-02637-z
Jouanin A, Borm T, Boyd LA, Cockram J, Leigh F, Santos BA, Visser RGF, Smulders MJM (2019a) Development of the GlutEnSeq capture system for sequencing gluten gene families in hexaploid bread wheat with deletions or mutations induced by γ-irradiation or CRISPR/Cas9. J Cereal Sci 88:157–166. https://doi.org/10.1016/j.jcs.2019.04.008
Jouanin A, Schaart JG, Boyd LA, Cockram J, Leigh FJ, Bates R, Wallington EJ, Visser RGF, Smulders MJM (2019b) Outlook for coeliac disease patients: towards bread wheat with hypoimmunogenic gluten by gene editing of α- and γ-gliadin gene families. BMC Plant Biol 19:333. https://doi.org/10.1186/s12870-019-1889-5
Jouanin A, Tenorio-Berrio R, Schaart JG, Zhang HQ, Yang ZJ, Yan BJ, Zhang HY (2020) Optimisation of droplet digital PCR for determining copy number variation of α-gliadin genes in mutant and gene-edited polyploid bread wheat. J Cereal Sci 9:102903. https://doi.org/10.1016/j.jcs.2019.102903
Kim MJ, Kim JK, Kim HJ, Pak JH, Lee JH, Kim DH, Kim HK, Jung HW, Lee JD, Chung YS, Ha SH (2012) Genetic modification of the soybean to enhance the β-carotene content through seed-specific expression. PLoS One 7:e48287. https://doi.org/10.1371/journal.pone.0048287
Lee JY, Kang CS, Beom HR, Jang YR, Altenbach SB, Lim SH, Kim YM, Park CS (2017) Characterization of a wheat mutant missing low-molecular-weight glutenin subunits encoded by the B-genome. J Cereal Sci 73:158–164. https://doi.org/10.1016/j.jcs.2016.12.004
Li D, Jin H, Zhang K, Wang Z, Wang F, Zhao Y, Huo N, Liu X, Gu YQ, Wang D, Dong L (2018) Analysis of the Gli-D2 locus identifies a genetic target for simultaneously improving the breadmaking and health-related traits of common wheat. Plant J 95:414–426. https://doi.org/10.1111/tpj.13956
Lim SH, Kim JK, Lee JY, KimYM SSH, Kim DH, Ha SH (2013) Petal-specific activity of the promoter of an anthocyanidin synthase gene of tobacco (Nicotiana tabacum L.). Plant Cell Tiss Organ Cult 114:373–383. https://doi.org/10.1007/s11240-013-0332-0
Morita E, Matsuo H, Mihara S, Morimoto K, Savage AW, Tatham AS (2003) Fast omega-gliadin is a major allergen in wheat-dependent exercise-induced anaphylaxis. J Dermatol Sci 33:99–104. https://doi.org/10.1016/s0923-1811(03)00156-7
Sánchez-León S, Gil-Humanes J, Ozuna CV, Giménez MJ, Sousa C, Voytas DF, Barro F (2018) Low-gluten, nontransgenic wheat engineered with CRISPR/Cas9. Plant Biotechnol J 16:902–910. https://doi.org/10.1111/pbi.12837
Schopf M, Scherf KA (2020) Predicting vital wheat gluten quality using the gluten aggregation test and the microscale extension test. Curr Res Food Sci 3:322–328. https://doi.org/10.1016/j.crfs.2020.11.004
Shimizu Y, Nasuda S, Endo TR (1997) Detection of the Sec-1 locus of rye by a PCR-based method. Genes Genet Syst 72:197–203. https://doi.org/10.1266/ggs.72.197
Skoczowski A, Obtułowicz K, Czarnobilska E, Dyga W, Mazur M, Stawoska I, Waga J (2017) Antibody reactivity in patients with IgE-mediated wheat allergy to various subunits and fractions of gluten and non-gluten proteins from omega-gliadin-free wheat genotypes. Ann Agric Environ Med 24:229–236. https://doi.org/10.5604/12321966.1233572
Spielmeyer W, Lagudah S (2003) Homoeologous set of NBS-LRR genes located at leaf and stripe rust resistance loci on short arms of chromosome 1 of wheat. Funct Integr Genomics 3:86–90. https://doi.org/10.1007/s10142-002-0074-2
Sukumaran S, Dreisigacker S, Lopes M, Chavez P, Reynolds MP (2015) Genome-wide association study for grain yield and related traits in an elite spring wheat population grown in temperate irrigated environments. Theor Appl Genet 128:353–363. https://doi.org/10.1007/s00122-014-2435-3
Tang ZX, Fu SL, Ren ZL, Zhang HQ, Yang ZJ, Yan BJ, Zhang HY (2008) Production of a new wheat cultivar with a different 1B.1R translocation with resistance to powdery mildew and stripe rust. Cereal Res Commun 36:451–460
Waga J, Skoczowski A (2014) Development and characteristics of omega-gliadin-free wheat genotypes. Euphytica 195:105–116. https://doi.org/10.1007/s10681-013-0984-1
Waga J, Zientarski J, Szaleniec M, Obtulowicz K, Dyga W, Skoczowski A (2013) Null alleles in gliadin coding loci and wheat allergenic properties. Am J Plant Sci 4:160–168. https://doi.org/10.4236/ajps.2013.41021
Wang DW, Li D, Wang J, Zhao Y, Wang Z, Yue G, Lin X, Ain H, Zhang K, Dong L, Wang D (2017) Genome-wide analysis of complex wheat gliadins, the dominant carriers of celiac disease epitopes. Sci Rep 7:44609. https://doi.org/10.1038/srep44609
Wen L, Tan B, Guo WW (2012) Estimating transgene copy number in precocious trifoliate orange by TapMan real-time PCR. Plant Cell Tissue Organ Cult 109:363–371. https://doi.org/10.1007/s11240-011-0101-x
Yahiaoui N, Srichumpa P, Dudler R, Keller B (2004) Genome analysis at different ploidy levels allows cloning of the powdery mildew resistance gene Pm3b from hexaploid wheat. Plant J 37:528–538. https://doi.org/10.1046/j.1365-313x.2003.0197
Yamamoto M, Mukai Y (2005) High-resolution physical mapping of the secalin-1 locus of rye on extended DNA fibers. Cytogenet Genome Res 109:79–82. https://doi.org/10.1159/000082385
Funding
This work was supported by grants from the Next‐Generation BioGreen 21 Linked Program (PJ015786) of the Rural Development Administration (RDA), Korea.
Author information
Authors and Affiliations
Contributions
Author contribution statement JYL designed the study and wrote the manuscript. SK conducted the experiments and wrote the manuscript. YQG, SBA, SDP and SHL reviewed the manuscript. JRS, FP and YJY performed the experiments. EJP and OT helped perform parts of experiments. Materials are supported by CSK and CC. All authors read and approved the final version of the paper.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing financial interests.
Ethics approval
Not applicable.
Additional information
Communicated by Lingrang Kong.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Kim, S., Sim, JR., Gu, Y.Q. et al. Toward reducing the immunogenic potential of wheat flour: identification and characterization of wheat lines missing omega-5 gliadins encoded by the 1D chromosome. Theor Appl Genet 136, 33 (2023). https://doi.org/10.1007/s00122-023-04295-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00122-023-04295-0