Abstract
Recently, electrophoretic analyses of waxy proteins in several hexaploid and tetraploid wheat accessions from worldwide collections have permitted the identification of new variants at the waxy loci, including allelic forms with different mobilities and partial null types. In this paper, the molecular characterization of mutated waxy loci in four bread wheat cultivars (two lacking the Wx-B1 and two lacking the Wx-D1 protein, respectively) and in four durum wheat cultivars (one lacking Wx-A1 and the remainder with Wx-B1 proteins showing different electrophoretic mobilities) was conducted by means of PCR, Southern and DNA sequence analyses. Three primer pairs were developed that identified six of the above-mentioned mutations and allowed their molecular description, providing a useful tool for further germplasm screening or marker assisted progeny selection in breeding programs involving the newly identified material. We have found that a complete gene deletion is responsible for a null allele at the Wx-B1 locus in one bread wheat line, whereas sequencing of the corresponding fragments showed a 724 bp deletion in the Wx-D1 locus in one line of bread wheat and an insertion of 89 bp in the Wx-A1 locus in one line of durum wheat, respectively. In addition, nucleotide substitutions and various insertions/deletions ranging from 3 to 30 bp were detected in the PCR fragments of one durum wheat line with a Wx-B1 protein with a different electrophoretic mobility. A fourth primer set, specific for this mutation, was consequently derived.
Similar content being viewed by others
References
Boggini G, Cattaneo M, Paganoni C, Vaccino P (2001) Genetic variation for waxy proteins and starch properties in Italian wheat germoplasm. Euphytica 119:111–114
Cai XL, Wang ZY, Xing YY, Zhang JL, Hong MM (1998) Aberrant splicing of intron 1 leads to the heterogeneous 5′ UTR and decreased expression of the Waxy gene in rice cultivars of intermediate amylose content. Plant J 14:459–465
Domon E, Fujita M, Ishikawa N (2002) The insertion/deletion polymorphism in the waxy gene of barley genetic resources from East Asia. Theor Appl Genet 104:132–138
D’Ovidio R, Tanzarella OA, Porceddu E (1992) Isolation of an alpha-type gliadin gene from T. durum Desf and genetic polymorphism at the Gli-2 loci. J Genet Breed 46:41–48
Dvorak J, McGuire PE, Cassidy B (1988) Apparent sources of the A genomes of wheats inferred from the polymorphism in abundance and restriction fragment length of repeated nucleotide sequences. Genome 30:680–689
Eriksson G (1970) The waxy character. Hereditas 63:180–204
Fedoroff N, Wessler S, Shure M (1983) Isolation of the transposable maize controlling elements Ac and Ds. Cell 85:235–242
Fredriksson H, Silverio J, Andersson R, Eliasson A-C, Åman P (1998) The influence of amylose and amylopectin characteristics on gelatinization and retrogradation properties of different starches. Carbohydr Polym 35:119–134
Graybosch RA, Peterson CJ, Hansen LE, Rahman S, Hill A, Skerritt JH (1998) Identification and characterization of US wheats carrying null alleles at the wx loci. Cereal Chem 75:162–165
Hirano HY, Eiguchi M, Sano Y (1998) A single base change altered the regulation of the waxy gene at the posttranscriptional level during the domestication of rice. Mol Biol Evol 15:978–987
Isshiki M, Morino K, Nakajima M, Okagaki RJ, Wessler SR, Izawa T, Shimamoto K (1998) A naturally occurring functional allele of the rice waxy locus has a GT to TT mutation at the 5′ splice site of the first intron. Plant J 15:133–138
James MG, Denyer K, Myers AM (2003) Starch synthesis in the cereal endosperm. Curr Opin Plant Biol 6:215–222
Lee M-R, Swanson BG, Baik B-K (2001) Influence of amylose content on properties of wheat starch and breadmaking quality of starch and gluten blends. Cereal Chem 78:701–706
Miura H, Tanii S (1994a) Endosperm starch properties in several wheat cultivars preferred for Japanese noodles. Euphytica 72:171–175
Miura H, Tanii S, Nakamura T, Watanabe N (1994b) Genetic control of amylose content in wheat endosperm starch and differential effects of three Wx genes. Theor Appl Genet 89:276–280
Miura H, Araki E, Tarui S (1999) Amylose synthesis capacity of the three Wx genes of wheat cv Chinese spring. Euphytica 108:91–95
Mohammadkhani A, Stoddard FL, Marshall DR, Uddin MN, Zhao X (1999) Starch extraction and amylose analysis from half seeds. Starch 51:62–66
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 234:71–79
Nakamura T, Yamamori M, Hirano H, Hidaka S (1993) Decrease of Waxy (Wx) protein in two common wheat cultivars with low amylose content. Plant Breed 111:99–105
Nakamura T, Yamamori M, Hirano H, Hidaka S, Nagamine T (1995) Production of waxy (amylose-free) wheats. Mol Gen Genet 248:253–259
Nakamura T, Vrinten P, Saito M, Konda M (2002) Rapid classification of partial waxy wheats using PCR-based markers. Genome 45:1150–1156
Nieto-Taladriz MT, Rodriguez-Quijano M, Carrillo JM (2000) Polymorphism of waxy proteins in Spanish durum wheats. Plant Breed 119:277–279
Patron NJ, Smith AM, Fahy BF, Hilton CM, Naldrett MJ, Rossnagel BG, Denyer K (2002) The altered pattern of amylose accumulation in the endosperm of low-amylose barley cultivars is attributable to a single mutant allele of granule-bound starch synthase I with a deletion in the 5′ non-coding region. Plant Physiol 130:190–198
Reddy I, Seib PA (2000) Modified waxy wheat starch copared to modified waxy corn starch. J Cereal Sci 31:25–39
Saito M, Konda M, Vrinten P, Nakamura K, Nakamura T (2004) Molecular comparison of waxy null alleles in common wheat and identification of a unique null allele. Theor Appl Genet 108:1205–1211
Urbano M, Margiotta B, Colaprico G, Lafiandra D (2002) Waxy protein in diploid, tetraploid and hexaploid wheats. Plant Breed 121:1–5
Vrinten P, Nakamura T, Yamamori M (1999) Molecular characterization of waxy mutations in wheat. Mol Gen Genet 261:463–471
Wessler SR, Varagona MJ (1985) Molecular basis of mutations at the waxy locus of maize: correlation with the fine structure genetic map. Proc Natl Acad Sci USA 82:4177–4182
Wessler SR, Baran G, Varagona MJ (1987) The maize transposable Ds element is spliced from RNA. Science 237:916–918
Yamamori M, Nakamura T, Endo TR, Nagamine T (1994) Waxy protein deficiency and chromosomal location of coding genes in common wheat. Theor Appl Genet 89:179–184
Yamamori M, Nakamura T, Nagamine T (1995) Polymorphism of two waxy proteins in the emmer group of tetraploid wheat, Triticum dicoccoides, T. dicoccum, and T. durum. Plant Breed 114:215–218
Yamamori M, Nakamura T, Kiribuchi-Otobe C (1998) Waxy protein alleles in common and emmer wheat germplasm. Misc Publ Natl Inst Agrobiol Resour 12:57–104
Yan L, Bhave M (2000) Sequence of the Waxy loci of wheat: utility in the analysis of waxy proteins and developing molecular markers. Biochem Genet 38:391–411
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 43:264–272
Yanagisawa T, Kiribuchi-Otobe C, Yoshida H (2001) An alanine to threonine change in the Wx-D1 protein reduces GBSS1 activity in waxy mutant wheat. Euphytica 121:209–214
Zao X C, Sharp PJ (1996) An improved 1-D SDS-PAGE method for the identification of three bread wheat waxy proteins. J Cereal Sci 23:191–193
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by G. Wenzel
Rights and permissions
About this article
Cite this article
Monari, A.M., Simeone, M.C., Urbano, M. et al. Molecular characterization of new waxy mutants identified in bread and durum wheat. Theor Appl Genet 110, 1481–1489 (2005). https://doi.org/10.1007/s00122-005-1983-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00122-005-1983-y