Development of a set of oligonucleotide primers specific for genes at the Glu-1 complex loci of wheat
1995
D'Ovidio, R. | Masci, S. | Porceddu, E. (Universita della Tuscia, Viterbo (Italy). Dept. di Agrobiologia e Agrochimica)
Specific amplification of the complete coding region of all six high-molecular-weight (HMW) glutenin genes present in hexaploid wheat was obtained by the polymerase chain reaction (PCR). Primers specific for the N-terminal region of the 1Dx gene and for the repetitive domain of the y-type HMW glutenin genes were also developed. Although the primers were constructed on the basis of the nucleotide sequences of HMW glutenin genes present in Triticum aestivum L. cv 'Cheyenne', they were very efficient in amplifying HMW glutenin genes of diploid and tetraploid wheat species. PCR analysis of HMW glutenin genes of Triticum urarto Tuman., Triticum longissimum Bowden and Triticum speltoides (Tausch) Gren. ex Richt. showed a high degree of length polymorphism, whereas a low degree of length variation was found in accessions of Triticum tauschii (Coss.) Schmal. Furthermore, using primers specific for the repetitive regions of HMW genes, it was possible to demonstrate that the size variation observed was due to a different length of the central repetitive domain. The usefulness of the PCR-based approach to analyze the genetic polymorphism of HMW glutenin genes, to isolate new allelic variants, to estimate their molecular size and to verify the number of cysteine residues is discussed.
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