Cloning and characterization of a gamma-3 hordein mRNA (cDNA) from Hordeum chilense (Roem. et Schult.)

Hordeum chilense is a wild relative of H. vulgare, cultivated barley, that has been successfully used in the synthesis of amphiploids by crossing with Triticum spp. These amphiploids-named generically xTritordeum-have been tested under field conditions, and one of them, the hexaploid tritordeum obtained following chromosome doubling of the hybrid H. chilense x T. turgidum, shows traits of interest inherited from the barley parent. Of great interest is the allelic variation observed in the endosperm storage proteins and their influence on the breadmaking and malting quality of tritordeum. We report here two mRNA (cDNA) sequences for a gamma-3 hordein from two accession lines of H. chilense, H1 and H7, and their characterization by quantitative real time (QRT)-PCR in the developing endosperm. Sequences were obtained by rapid amplification of cDNA ends and edge-to-edge amplification of open reading frames from cDNA of H. chilense. Eight putative single nucleotide polymorphisms and one codon insertion were identified in the sequences of the H1 and H7 gamma-3 hordeins. The deduced amino acid sequences showed similar features to that of the gamma-3 hordein and gamma-gliadins from barley and wheat, respectively. While the repetitive motif (PQQQPF) is similar to that of the gamma-3 hordein from H. vulgare, there are 19 motif repeats in H. vulgare, whereas H. chilense shows 15 tandem repeats. The transcription of the genes encoding for the gamma-3 hordein were monitored by QRT-PCR: in both lines maximum transcription occurred 12 days after flowering.