Molecular Breeding

, Volume 34, Issue 3, pp 1137–1146 | Cite as

Wx gene in diploid wheat: molecular characterization of five novel alleles from einkorn (Triticum monococcum L. ssp. monococcum) and T. urartu



The Wx gene encodes the granule-bound starch synthase I or waxy protein, which is the sole enzyme responsible for amylose synthesis in wheat seeds. Triticum urartu and einkorn (T. monococcum L. ssp. monococcum), which are related to the A genome of bread wheat, could be important sources of variation for this gene. This study evaluated the Wx gene variability in 52 accessions of these species and compared their nucleotide sequences with the Wx-A1a allele of bread wheat. The level of polymorphism found was high, although not distributed equally between the two species. Five different alleles were found in T. urartu, of which four were novel (Wx-A u 1b, -A u 1c, -A u 1d and -A u 1e). All einkorn accessions had the same allele, which was also novel and was named Wx-A m 1a. A comparison between the proteins deduced from the novel alleles and the Wx-A1a protein showed that there were up to 33 amino acid changes in both the transit peptide and the mature protein. These results showed that these species, especially T. urartu, are a potential source of novel waxy variants.


Einkorn Molecular characterization Starch T. urartu Wx gene 



This research was supported by grant AGL2010-19643-C02-01 from the Spanish Ministry of Economy and Competitiveness, co-financed with the European Regional Development Fund (FEDER) from the European Union. We thank the National Small Grain Collection (Aberdeen, USA) and the Institute for Plant Genetics and Crop Plant Research (Gatersleben, Germany) for supplying the material analyzed.

Supplementary material

11032_2014_105_MOESM1_ESM.pdf (172 kb)
Supplementary material 1 (PDF 172 kb)


  1. Ainsworth C, Clark J, Balsdon J (1993) Expression, organisation and structure of the genes encoding the waxy protein (granule-bound starch synthase) in wheat. Plant Mol Biol 22:67–82PubMedCrossRefGoogle Scholar
  2. Baldwin PM (2001) Starch granule-associated protein and polypeptides: a review. Starch/Stärcke 53:475–503CrossRefGoogle Scholar
  3. Brandolini A, Vaccino P, Boggini G, Özkan H, Kilian B, Salamini F (2006) Quantification of genetic relationships among A genomes of wheats. Genome 49:297–305PubMedCrossRefGoogle Scholar
  4. Caballero L, Bancel E, Debiton C, Branlard G (2008) Granule-bound starch synthase (GBSS) diversity of ancient wheat and related species. Plant Breed 127:548–553CrossRefGoogle Scholar
  5. Chao S, Sharp PJ, Worland AJ, Warham EJ, Koebner RMD, Gale MD (1989) RFLP-based genetic maps of wheat homoeologous group 7 chromosomes. Theor Appl Genet 78:495–504PubMedCrossRefGoogle Scholar
  6. Dvorak J, McGuire PE, Cassidy B (1988) Apparent source of the A genomes of wheats inferred from polymorphism in abundance and restriction fragment length of repeated nucleotide sequences. Genome 30:680–689CrossRefGoogle Scholar
  7. Dvorak J, Pd Terlizzi, Zhang H-B, Resta P (1993) The evolution of polyploid wheats: identification of the A genome donor species. Genome 36:21–31PubMedCrossRefGoogle Scholar
  8. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  9. Feuillet C, Langridge P, Waugh R (2008) Cereal breeding takes a walk on the wild side. Trends Genet 24:24–32PubMedCrossRefGoogle Scholar
  10. Guzmán C, Alvarez JB (2012) Molecular characterization of a novel waxy allele (Wx-A u 1a) from Triticum urartu Thum. ex Gandil. Genet Resour Crop Evol 59:971–979CrossRefGoogle Scholar
  11. Guzmán C, Caballero L, Alvarez JB (2009) Variation in Spanish cultivated einkorn wheat (Triticum monococcum L. ssp. monococcum) as determined by morphological traits and waxy proteins. Genet Resour Crop Evol 56:601–604CrossRefGoogle Scholar
  12. Guzmán C, Caballero L, Moral A, Alvarez JB (2010) Genetic variation for waxy proteins and amylose content in Spanish spelt wheat (Triticum spelta L.). Genet Resour Crop Evol 57:721–725CrossRefGoogle Scholar
  13. Guzmán C, Caballero L, Alvarez JB (2011) Molecular characterisation of the Wx-B1 allelic variants identified in cultivated emmer wheat and comparison with those of durum wheat. Mol Breed 28:403–411CrossRefGoogle Scholar
  14. Guzmán C, Caballero L, Martín LM, Alvarez JB (2012) Waxy genes from spelt wheat: new alleles for modern wheat breeding and new phylogenetic inferences about the origin of this species. Ann Bot 110:1161–1171PubMedCrossRefPubMedCentralGoogle Scholar
  15. Huang XQ, Brûlé-Babel A (2012) Sequence diversity, haplotype analysis, association mapping and functional marker development in the waxy and starch synthase IIa genes for grain-yield-related traits in hexaploid wheat (Triticum aestivum L.). Mol Breed 30:627–635CrossRefGoogle Scholar
  16. Kiribuchi-Otobe C, Nagamine T, Yanagisawa T, Ohnishi M, Yamaguchi I (1997) Production of hexaploid wheats with waxy endosperm character. Cereal Chem 74:72–74CrossRefGoogle Scholar
  17. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  18. Leterrier M, Holappa LD, Broglie KE, Beckles DM (2008) Cloning, characterisation and comparative analysis of a starch synthase IV gene in wheat: functional and evolutionary implications. BMC Plant Biol 8:98PubMedCrossRefPubMedCentralGoogle Scholar
  19. Li W, Gao Z, Xiao W, Wei YM, Liu YX, Chen GY, Pu ZE, Chen HP, Zheng YL (2012) Molecular diversity of restriction enzyme sites, Indels and upstream open reading frames (uORFs) of 5′ untranslated regions (UTRs) of Waxy genes in Triticum L. and Aegilops L. species. Genet Resour Crop Evol 59:1625–1647CrossRefGoogle Scholar
  20. Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452PubMedCrossRefGoogle Scholar
  21. Liu YX, Li W, Wei YM, Chen GY, Zheng YL (2009) Molecular characterization of the waxy gene in einkorn wheat. J Plant Sci 4:114–121CrossRefGoogle Scholar
  22. Mason-Gamer RJ, Weil CF, Kellogg EA (1998) Granule-bound starch synthase: structure, function, and phylogenetic utility. Mol Biol Evol 15:1658–1673PubMedCrossRefGoogle Scholar
  23. McIntosh RA, Yamazaki Y, Dubcovsky J, Rogers WJ, Morris C, Appels R, Xia XC (2013) Catalogue of gene symbols for wheat. Accessed 22 Apr 2014
  24. Miller TE, Reader SM (1980) Variation in the meiotic chromosome pairing of hybrids between hexaploid and diploid wheats. Cereal Res Commun 8:477–483Google Scholar
  25. Miura H, Sugawara A (1996) Dosage effects of the three Wx genes on amylose synthesis in wheat endosperm. Theor Appl Genet 93:1066–1070PubMedCrossRefGoogle Scholar
  26. Monari AM, Simeone MC, Urbano M, Margiotta B, Lafiandra D (2005) Molecular characterization of new waxy mutants identified in bread and durum wheat. Theor Appl Genet 110:1481–1489PubMedCrossRefGoogle Scholar
  27. Murai J, Taira T, Ohta D (1999) Isolation and characterization of the three Waxy genes encoding the granule-bound starch synthase in hexaploid wheat. Gene 234:71–79PubMedCrossRefGoogle Scholar
  28. Nakamura T, Yamamori M, Hirano H, Hidaka S, Nagamine T (1995) Production of waxy (amylose-free) wheats. Mol Gen Genet 248:253–259PubMedCrossRefGoogle Scholar
  29. Nei M (1987) Molecular evolutionary genetics. Columbia University Press, New YorkGoogle Scholar
  30. Ortega R, Alvarez JB, Guzmán C (2014) Characterization of the Wx gene in diploid Aegilops species and its potential use in wheat breeding. Genet Resour Crop Evol 61:369–382CrossRefGoogle Scholar
  31. Rawat N, Sehgal SK, Joshi A, Rothe N, Wilson DL, McGraw N, Vadlani PV, Li W, Gill BS (2012) A diploid wheat TILLING resource for wheat functional genomics. BMC Plant Biol 12:205PubMedCrossRefPubMedCentralGoogle Scholar
  32. Rice P, Longden I, Bleasby A (2000) EMBOSS: the European molecular biology open software suite. Trends Genet 16:276–277PubMedCrossRefGoogle Scholar
  33. Rodriguez-Quijano M, Vázquez JF, Carrillo JM (2004) Waxy proteins and amylose content in diploid Triticeae species with genomes A, S and D. Plant Breed 123:294–296CrossRefGoogle Scholar
  34. Sim NL, Kumar P, Hu J, Henikoff S, Schneider G, Ng PC (2012) SIFT web server: predicting effects of amino acid substitutions on proteins. Nucleic Acids Res 40(W1):W452–W457PubMedCrossRefPubMedCentralGoogle Scholar
  35. Slade AJ, Fuerstenberg SI, Loeffler D, Steine MN, Facciotti D (2005) A reverse genetic, nontransgenic approach to wheat crop improvement by TILLING. Nat Biotechnol 23:75–81PubMedCrossRefGoogle Scholar
  36. Stacey J, Isaac P (1994) Isolation of DNA from plants. In: Isaac PG (ed) Methods in molecular biology: protocols for nucleic acid analysis by non-radiactive probes. Humana Press, Totawa, pp 9–15Google Scholar
  37. Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci USA 101:11030–11035PubMedCrossRefPubMedCentralGoogle Scholar
  38. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739PubMedCrossRefPubMedCentralGoogle Scholar
  39. Urbano M, Margiotta B, Colaprico G, Lafiandra D (2002) Waxy proteins in diploid, tetraploid and hexaploid wheats. Plant Breed 121:465–469CrossRefGoogle Scholar
  40. Watterson GA (1975) On the number of segregating sites in genetical models without recombination. Theor Popul Biol 7:256–276PubMedCrossRefGoogle Scholar
  41. Wu Y, Weber JL, Vladutiu GD, Tarnopolsky MA (2011) Six novel mutations in the myophosphorylase gene in patients with McArdle disease and a family with pseudo-dominant inheritance pattern. Mol Genet Metab 104:587–591PubMedCrossRefGoogle Scholar
  42. Yamamori M (2009) Amylose content and starch properties generated by five variant Wx alleles for granule-bound starch synthase in common wheat (Triticum aestivum L.). Euphytica 165:607–614CrossRefGoogle Scholar
  43. Yamamori M, Guzmán C (2013) SNPs and an insertion sequence in five Wx-A1 alleles as factors for variant Wx-A1 protein in wheat. Euphytica 192:325–338CrossRefGoogle Scholar
  44. Yamamori M, Yamamoto K (2011) Effects of two novel Wx-A1 alleles of common wheat (Triticum aestivum L.) on amylose and starch properties. J Cereal Sci 54:229–235CrossRefGoogle Scholar
  45. Yamamori M, Nakamura T, Endo TR, Nagamine T (1994) Waxy protein deficiency and chromosomal location of coding genes in common wheat. Theor Appl Genet 89:179–184PubMedCrossRefGoogle Scholar
  46. Yamamori M, Fujita S, Hayakawa K, Matsuki J, Yasui T (2000) Genetic elimination of a starch granule protein, SGP-1, of wheat generates an altered starch with apparent high amylose. Theor Appl Genet 101:21–29CrossRefGoogle Scholar
  47. Yan L, Bhave M (2001) Characterization of waxy proteins and waxy genes of Triticum timopheevii and T. zhukovskyi and implications for evolution of wheat. Genome 44:582–588PubMedCrossRefGoogle Scholar
  48. Yan L, Bhave M, Fairclough R, Konik C, Rahman S, Appels R (2000) The genes encoding granule-bound starch synthases at the waxy loci of the A, B, and D progenitors of common wheat. Genome 43:264–272PubMedCrossRefGoogle Scholar
  49. Yasui T, Sasaki T, Matsuki J, Yamamori M (1997) Waxy endosperm mutants of bread wheat (Triticum aestivum L.) and their starch properties. Breed Sci 47:161–163Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Raúl Ortega
    • 1
  • Carlos Guzmán
    • 1
  • Juan B. Alvarez
    • 1
  1. 1.Departamento de Genética, Escuela Técnica Superior de Ingeniería Agronómica y de Montes, Edificio Gregor Mendel, Campus de RabanalesUniversidad de Córdoba, CeiA3CórdobaSpain

Personalised recommendations