Abstract
Allelic variations in storage-protein genes play a significant role in determining the properties of wheat flour. This study aimed to investigate allelic diversity in gene blocks encoding seed storage proteins in tetraploid wheat, specifically α/β-gliadin and low-molecular-weight glutenin subunits (LMW-GS). DNA was extracted from 90 rain-fed durum wheat genotypes, followed by a polymerase chain reaction (PCR) with 12 specific primers targeting Gli-A2, Gli-B2, Glu-A3, and Glu-B3 gene loci. The PCR products were then analyzed on a 2% agarose gel. For the Gli-A2 gene block, three pairs of specific primer identified a total of 13 alleles, while the Gli-B2 gene block revealed two alleles using the gli-AS_2 primer. In the Glu-A3 gene block, six primer pairs detected 42 alleles, and the Glu-B3 gene block showed eight alleles using 2 primer pairs. In total, we identified 7 major alleles and 8 minor alleles at Gli-2 loci, and 12 major alleles and 38 minor alleles at Glu-3 loci. Different primers yielded varying banding patterns, with the highest alleles and banding pattern counts observed with Glu3A.1 primer. Genetic dissimilarities ranged from 0.030 (between G21 and G83) to 0.583 (between G4 and G51). Structure analysis categorized the durum wheat collection into five subpopulations. Results confirmed a substantial variation at the Gli-2 and Glu-3 loci in tetraploid wheat.
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Notes
The negative sign indicates the absence, and the positive sign indicates the presence of the allele. The numbers above the symbols indicate the size of the allele in base pairs.
References
Ahmadi J, Pour-Aboughadareh A (2015) Allelic variation of glutenin and gliadin genes in Iranian einkorn wheat. J Biodivers Environ Sci 7:168–179
Al-Khayri JM, Alshegaihi RM, Mahgoub EI, Mansour E, Atallah OO, Sattar MN, Al-Mssallem MQ, Alessa FM, Aldaej MI, Hassanin AA (2023) Association of high and low molecular weight glutenin subunits with gluten strength in tetraploid durum wheat (Triticum turgidum spp. Durum L.). Plants 12:1416. https://doi.org/10.3390/plants12061416
An X, Zhang Q, Yan Y, Li Q, Zhang Y, Wang A, Pei Y, Tian J, Wang H, Hsam SLK, Zeller FJ (2006) Cloning and molecular characterization of three novel LMW-i glutenin subunit genes from cultivated einkorn (Triticum monococcum L.). Theor Appl Genet 113:383–395. https://doi.org/10.1007/s00122-006-0299-x
Boehm JD, Ibba MI, Kiszonas AM, Morris CF (2017) End-use quality of CIMMYT-derived soft-kernel durum wheat Germplasm: II. dough strength and pan bread quality. Crop Sci 57:1485–1494. https://doi.org/10.2135/cropsci2016.09.0775
Carvalho MA, Quesenberry KH, Gallo M (2010) Comparative assessment of variation in the USA Arachis pintoi (Krap. and Greg.) germplasm collection using RAPD profiling and tissue culture regeneration ability. Plant Syst Evol 288:245–251. https://doi.org/10.1007/s00606-010-0335-9
Chegdali Y, Ouabbou H, Essamadi A, Cervantes F, Ibba MI, Guzmán C (2020) Assessment of the glutenin subunits diversity in a durum wheat (T. turgidum ssp. durum) collection from Morocco. Agronomy 10:957. https://doi.org/10.3390/agronomy10070957
Chen C, Chu Y, Ding C, Su X, Huang Q (2020) Genetic diversity and population structure of black cottonwood (Populus deltoides) revealed using simple sequence repeat markers. BMC Genet 21:2. https://doi.org/10.1186/s12863-019-0805-1
Dai S, Xu D, Yan Y, Wen Z, Zhang J, Chen H, Lu Z, Li H, Cong H, Wei Y, Zheng Y, Yan Z (2020) Characterization of high- and low-molecular-weight glutenin subunits from Chinese **njiang wheat landraces and historical varieties. J Food Sci Technol 57:3823–3835. https://doi.org/10.1007/s13197-020-04414-5
Delcour JA, Joye IJ, Pareyt B, Wilderjans E, Brijs K, Lagrain B (2012) Wheat gluten functionality as a quality determinant in cereal-based food products. Annu Rev Food Sci Technol 3:469–492. https://doi.org/10.1146/annurev-food-022811-101303
D’Ovidio R, Masci S (2004) The low-molecular-weight glutenin subunits of wheat gluten. J Careal Sci 39:321–339. https://doi.org/10.1016/j.jcs.2003.12.002
Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conserv Genet Resour 4:359–361. https://doi.org/10.1007/s12686-011-9548-7
El Haddad N, Kabbaj H, Zaïm M, El Hassouni K, TidianeSall A, Azouz M, Ortiz R, Baum M, Amri A, Gamba F, Bassi FM (2021) Crop wild relatives in durum wheat breeding: drift or thrift? Crop Sci 61:37–54. https://doi.org/10.1002/csc2.20223
Etminan A, Pour-Aboughadareh A, Mohammadi R, Noori A, Ahmadi-Rad A (2018) Applicability of CAAT Box-derived Polymorphism (CBDP) markers for analysis of genetic diversity in durum wheat. Cereal Res Commun 46:1–9. https://doi.org/10.1556/0806.45.2017.054
Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620. https://doi.org/10.1111/j.1365-294X.2005.02553.x
Henkrar F, El-Haddoury J, Ouabbou H, Nsarellah N, Iraqi D, Bendaou N, Udupa SM (2016) Genetic diversity reduction in improved durum wheat cultivars of Morocco as revealed by microsatellite markers. Sci Agric 73:134–141. https://doi.org/10.1590/0103-9016-2015-0054
Hsia CC, Anderson OD (2001) Isolation and characterization of wheat ω-gliadin genes. Theor Appl Genet 103:37–44. https://doi.org/10.1007/s00122-001-0552-2
Hu X, Peng Y, Ren X, Peng J, Nevo E, Ma W, Sun D (2018) Allelic variation of low molecular weight glutenin subunits composition and the revealed genetic diversity in durum wheat (Triticum turgidum L. ssp. durum (Desf)). Breed Sci 68:524–535. https://doi.org/10.1270/jsbbs.18085
Huo N, Dong L, Zhang S, Wang Y, Zhu T, Mohr T, Altenbach S, Liu Z, Dvorak J, Anderson OD, Luo M-C, Wang D, Gu YQ (2017) New insights into structural organization and gene duplication in a 1.75-Mb genomic region harboring the α-gliadin gene family in Aegilops tauschii, the source of wheat D genome. Plant J 92:571–583. https://doi.org/10.1111/tpj.13675
Irani P (2000) Pasta quality traits of some durum wheat varieties. J Agric Sci Technol 2:143–148
Kawaura K, Mochida K, Ogihara Y (2006) Expression profile of two storage-protein gene families in hexaploid wheat revealed by large-scale analysis of expressed sequence tags. Plant Physiol 139:1870–1880. https://doi.org/10.1104/pp.105.070722
Long H, Wei Y-M, Yan Z-H, Baum B, Nevo E, Zheng Y-L (2005) Classification of wheat low-molecular-weight glutenin subunit genes and its chromosome assignment by develo** LMW-GS group-specific primers. Theor Appl Genet 111:1251–1259. https://doi.org/10.1007/s00122-005-0024-1
Long H, Huang Z, Wei YM, Yan ZH, Ma ZC, Zheng YL (2008) Length variation of i-type low-molecular-weight glutenin subunit genes in diploid wheats. Russ J Genet 44:429–435. https://doi.org/10.1134/S102279540804008X
Mohammadi R, Haghparast R (2022) Durum wheat: production, nutritional value and economic importance. Cereal Biotechnol Biochem 1:414–445. https://doi.org/10.22126/cbb.2022.8385.1021
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325. https://doi.org/10.1093/nar/8.19.4321
Negisho K, Shibru S, Pillen K, Ordon F, Wehner G (2021) Genetic diversity of Ethiopian durum wheat landraces. PLoS ONE 16:e0247016. https://doi.org/10.1371/journal.pone.0247016
Nigro D, Monaci L, Pilolli R, Gadaleta A (2020) Allelic variation of gliadin-encoding genes in a collection of tetraploid wheat genotypes. Cereal Res Commun 48:507–515. https://doi.org/10.1007/s42976-020-00061-w
Noma S, Hayakawa K, Abe C, Suzuki S, Kawaura K (2019) Contribution of α-gliadin alleles to the extensibility of flour dough in Japanese wheat cultivars. J Careal Sci 86:15–21. https://doi.org/10.1016/j.jcs.2018.12.017
Noma S, Yamagishi M, Ogihara Y, Kawaura K (2023) Characterization of α-gliadin alleles of Japanese wheat cultivars in relation to flour dough extensibility and celiac disease epitopes. J Careal Sci 109:103591. https://doi.org/10.1016/j.jcs.2022.103591
Nour-Mohammadi E, Fabriki-Ourang S, Pour-Abughadareh A (2021) Evaluation of allelic diversity of genes encoding glutenin and gliadin in bread wheat and its some relatives. Mod Genet 16:329–340
Osborne TB (1907) The proteins of the wheat kernel. Carnegie Institution of Washington, Washington, D.C.
Payne PI (1987) Genetics of wheat storage proteins and the effect of allelic variation on bread-making quality. Annu Rev Plant Physiol 38:141–153. https://doi.org/10.1146/annurev.pp.38.060187.001041
Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics 28:2537–2539. https://doi.org/10.1093/bioinformatics/bts460
Pei Y-H, Wang A-L, An X-L, Li X-H, Zhang Y-Z, Huang X-Q, Yan Y-M (2007) Characterization and comparative analysis of three low molecular weight glutenin C-subunit genes isolated from Aegilops tauschii. Can J Plant Sci 87:273–280. https://doi.org/10.4141/p06-152
Peña E, Bernardo A, Soler C, Jouve N (2005) Relationship between common wheat (Triticum aestivum L.) gluten proteins and dough rheological properties. Euphytica 143:169–177. https://doi.org/10.1007/s10681-005-3157-z
Que Y, Pan Y, Lu Y, Yang C, Yang Y, Huang N, Xu L (2014) Genetic analysis of diversity within a chinese local sugarcane germplasm based on start codon targeted polymorphism. Biomed Res Int 2014:468375. https://doi.org/10.1155/2014/468375
Shariat F, Mohammadi SA, Norouzi M, Valizadeh M (2016) Allelic diversity of low-molecular-weight glutenin subunit genes in Iranian winter wheat landrace. Careal Res 6:65–77
Shaygan N, Etminan A, MajidiHervan I, Azizinezhad R, Mohammadi R (2021) The study of genetic diversity in a minicore collection of durum wheat genotypes using agro-morphological traits and molecular markers. Cereal Res Commun 49:141–147. https://doi.org/10.1007/s42976-020-00073-6
Wang L, Li G, Peña RJ, **a X, He Z (2010) Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.). J Cereal Sci 51:305–312. https://doi.org/10.1016/j.jcs.2010.01.005
Yang FP, Wang LH, Wang JW, He XY, Zhang XK, Shang XW, Yang WX, **a XC, He ZH (2009) 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. Crop Pasture Sci 61:84–91. https://doi.org/10.1071/CP09164
Zang R, Zhao Y, Guo K, Hong K, ** H, Wen C (2018) The population genetic variation analysis of bitter gourd wilt caused by Fusarium oxysporum f. sp. momordicae in China by inter simple sequence repeats (ISSR) molecular marker. bioRxiv 424077. https://doi.org/10.1101/424077
Zhang W, Gianibelli MC, Rampling LR, Gale KR (2004) Characterisation and marker development for low molecular weight glutenin genes from Glu-A3 alleles of bread wheat (Triticum aestivum. L). Theor Appl Genet 108:1409–1419. https://doi.org/10.1007/s00122-003-1558-8
Zhen S, Han C, Ma C, Gu A, Zhang M, Shen X, Li X, Yan Y (2014) Deletion of the low-molecular-weight glutenin subunit allele Glu-A3a of wheat (Triticum aestivum L.) significantly reduces dough strength and breadmaking quality. BMC Plant Biol 14:367. https://doi.org/10.1186/s12870-014-0367-3
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The authors would like to thank the Sararud Dryland Agricultural Research Institute of Kermanshah for its sincere cooperation in providing the genotypes.
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Moradi Kheibari, Z., Azizinezhad, R., Mehrabi, A. et al. Allelic variation and genetic diversity of low-molecular-weight glutenin subunit- and α/β-gliadin encoding genes in tetraploid wheat. CEREAL RESEARCH COMMUNICATIONS (2024). https://doi.org/10.1007/s42976-024-00527-1
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DOI: https://doi.org/10.1007/s42976-024-00527-1