Biopolym. Cell. 2021; 37(5):379-388.
Genomics, Transcriptomics and Proteomics
Genetic variation of GLI-B1 locus in ukrainian bread wheat varieties and lines
1Popovych Yu. A., 2Blagodarova O. M., 1, 2Chebotar S. V.
  1. Odessa I. I. Mechnikov National University
    2, Dvoryanskaya Str., Odessa, Ukraine, 65082
  2. Plant breeding and Genetics Institute, National Centre of Seed and Cultivar Investigation of NAAS
    Ovidiopol'skaya dor., 3, Odessa, Ukraine, 65036


Aim. To investigate polymorphism of Gli-B1 locus in modern Ukrainian bread wheat cultivars, to analyze the distribution of alleles and to compare received data with“core-collection of wheat cultivars” presented by Dr. E. Metakovsky. Methods. Eighty one bread wheat cultivars and lines from different plant breeding institutions and stations of Ukraine were tested using allele-specific primers to Gli-B1 locus developed by Zhang et al. [2003]. PCR products were fractionated in polyacrylamide gel (PAG) and then were stained by silver nitrate. Allelic variants of gliadins were analyzed by electrophoresis in acid polyacrylamide gel (APAGE). Results. Nine allelic variants of gliadins were revealed by APAGE and six alleles of Gli-B1 locus were detected by PCR-analysis. In 52 % of modern Ukrainian bread wheat cultivars we revealed Gli-B1b allelic variant, according to PCR - Gli-B1.1 allele with a 360-bp amplification fragment. In the genotypes of Ukrainian wheat cultivars, the 1RS.1BL translocation, which carries resistance genes, is frequent, as was detected by the absence of any amplification fragments with Gli-B1 primers. The correspondence between allelic variants of gliadins and alleles of Gli-B1 locus is discussed. Conclusions. DNA polymorphism of Gli-B1 locus examined in our research coincides with the diversity of allelic variants of gliadins, which were detected by APAGE method for Ukrainian bread wheat cultivar. However, PCR-analysis with applied primers carried out in this study does not distinguish the alleles that correspond to the Gli-B1c, Gli-B1g and Gli-B1e allelic variants of gliadins. The most common allele (52 %) for the investigated Ukrainian wheat varieties is Gli-B1.1 allele, which was characterized by the amplification fragment of 369 bp, and the presence of 1RS.1BL translocation, which corresponds to Gli-B1b and Gli-B1l allelic variants of gliadins obtained by APAGE, respectively.
Keywords: Triticum aestivum L., Gli-B1, gliadins, polymorphism, PCR analysis


[1] Urade R, Sato N, Sugiyama M. Gliadins from wheat grain: an overview, from primary structure to nanostructures of aggregates. Biophys Rev. 2018; 10:435-43.
[2] Gianibelli MC, Larroque OR, MacRitchie F, Wrigley CW. Biochemical, genetic, and molecular characterization of wheat glutenin and its component subunits. Cereal Chem. 2001; 78(6):635-46.
[3] Wieser H. Chemistry of gluten proteins. Food Microbiol. 2007; 24(2):115-9.
[4] Vriezinga SL, Schweizer JJ, Koning F, Mearin ML. Coeliac disease and glutenrelated disorders in childhood. Nat Rev Gastroenterol Hepatol. 2015; 12:527-36.
[5] Catassi C, Fasano A. Coeliac disease: The debate on coeliac disease screening - are we there yet? Nat Rev Gastroenterol Hepatol. 2014; 11:457-8.
[6] Wang DW, Li D, Wang J, Zhao Y, Wang Zh, Yue GLX, Huanju Q, Zhang K, Dong L, Wang D. Genome-wide analysis of complex wheat gliadins, the dominant carriers of celiac disease epitopes. Sci Rep. 2017; 7:44609.
[7] Jouanin A, Schaart JG, Boyd LA, et al. Outlook for coeliac disease patients: towards bread wheat with hypoimmunogenic gluten by gene editing of α- and γ-gliadin gene families. BMC Plant Biol. 2019; 19: 333.
[8] Li D, Jin H, Zhang K, Wang Zh, Wang F, Zhao Y, Huo N, Liu X, Gu Y, Wang D, Dong L. Analysis of the Gli-D2 locus identifies a genetic target for simultaneously improving the breadmaking and health-related traits of com-mon wheat. Plant J. 2018; 95(3):414-426.
[9] Wrigley CW, Shepherd KW. Electrofocusing of grain proteins from wheat genotypes. Ann NY Acad Sci. 1973; 209(1):154-62.
[10] Anderson OD, Dong L, Huo N, Gu, YQ. A new class of wheat gliadin genes and proteins. PLoS One. 2012; 7:e52139
[11] Payne PI, Holt LM, Lawrence GJ, Law CN. The genetics of gliadin and glutenin, the major storage proteins of the wheat endosperm. Plant Food Hum Nutr. 1982; 31:229-41.
[12] Hafeez AN, Arora S, Ghosh S, Gilbert D, Bowden R L, Wulff BH. Creation and judicious application of a wheat resistance genes atlas. Mol Plant. 2021; 14(7):1053-70.
[13] Huang L, Brooks SA, Li W, Fellers JP, Trick HN, Gill BS. Map-based cloning of leaf rust resistance gene Lr21 from the large and polyploid genome of bread wheat. Genetics. 2003; 164(2): 655-64.
[14] Anderson OD, Greene FC. The α-gliadin gene family. II DNA and protein sequence variation, subfamily structure, and origins of pseudogenes. Theor Appl Genet. 1997; 95: 59-65.
[15] Okita TW, Cheesbrough V, Reeves CD. Evolution and heterogeneity of the α-/β-type and γ-type gliadin DNA sequences. J Biol Chem. 1985; 260(13):8203-13.
[16] Hsia CC, Anderson OD. Isolation and characterization of wheat gliadin genes. Theor Appl Genet. 2001; 103: 37-44.
[17] Sozinov AA, Poperelya FA. Genetic classification of prolamins and its use for plant breeding. Ann Technol Agric. 1980; 29(2):229-45.
[18] Metakovsky E, Melnik VA, Rodriguez-Quijano M, Upelniek VP, Carrillo JM. A catalog of glidin alleles: Poly-morphism of 20th-century common wheat germplasm. Crop J. 2018; 6(6):629-41.
[19] Sozinov AA, Poperelya FA. Prolamin polymorphism and breeding. Visn sel-khozh nayki. 1979; 10:21-34.
[20] Huo N, Zhang S, Zhu T, Dong L, Wang Y, Mohr T, Hu T, Liu Z, Dvorak J, Luo MC, Wang D, Lee JY, Altenbach S, Gu YQ. Gene duplication and evolution dynamics in the homeologous regions harboring multiple prolamin and resistance gene families in hexaploid wheat. Front Plant Sci. 2018; 9:673.
[21] Zhang, Gianibelli M, Rampling ML, Gale KR. Identification of SNPs and development of allele-specific PCR markers for γ-gliadin alleles in Triticum aestivum. Theor Appl Genet. 2003; 107: 130-8.
[22] Devos KM, Bryan GJ, Collins AJ, Stephenson P, Gale MD. Application of two microsatellite sequences in wheat storage proteins as molecular markers. Theor Appl Genet. 1995; 90:247-52.
[23] Polischuk AM, Chebotar SV, Blagodarova OM, Kozub NA, Sozinov IA, Sivolap YuM. Analysis of varieties and near-isogenic lines of bread wheat by PCR with allele-specific primers to Gli-1 and Glu-3 loci. Cytol Genet. 2010; 44:345-53.
[24] Popovych Y, Chebotar S, Melnik V, Rodriguez-Quijano M, Pascual L, Rogers WJ, Metakovsky E. Congruity of the polymorphisms in the expressed and noncoding parts of the Gli-B1 locus in common wheat. Agronomy. 2020; 10(10):1510.
[25] Copus MM. About the natural gene geography of gliadin alleles in winter bread wheat. Selektsya & semenovodstvo. 1994; 5:9-14.
[26] Poperelya FA. Gliadin polymorphism and its association with grain quality, productivity and adaptation properties of winter bread wheat varieties. Breeding, seed production and intensive technology of wheat cultivation. Moscow: Agropromizdat. 1989:138-50.
[27] Doyle JJ, Doyle JL. Isolation of plant DNA from fresh tissue. Focus. 1990; 12:13-15.
[28] Promega Technical Manual. Gene Print. STR Systems. Printed in USA. Revised. 1999; 7:52.
[29] Kozub NA, Sozinov IA, Sobko TA, Kolyuchii VT, Kuptsov SV, Sozinov AA. Variation at storage protein loci in winter common wheat cultivars of the central forest-steppe of Ukraine. Cytol Genet. 2009; 43:55-62.
[30] Nei M. Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci USA. 1973; 70(12): 3321-3.
[31] Kozub NO, Sozinov IO, Chaika VM, Sozinova OI, Janse LA, Blume YaB. Changes in allele frequencies at storage protein loci of winter common wheat under climate change. Cytol Genet. 2020; 54:305-17.
[32] Blagodarova OM, Lytvynenko MA, Golub YeA. Gene geography of alleles at gliadin- and glutenin-coding loci of Ukrainian winter common wheat varieties and their association with agronomical traits. Coll Sci Pap Inst Breed Genet. 2004; 46(6):124-38.
[33] Kozub NA, Sozinov IA, Karelov AV, Blume YaB, Sozinov AA. Diversity of Ukrainian winter common wheat varieties with respect to storage protein loci and molecular markers for disease resistance genes. Cytol Genet. 2017; 51:117-129.