The development of a PCR-based marker for PSY1 from Hordeum chilense, a candidate gene for carotenoid content accumulation in tritordeum seeds

Hexaploid tritordeums are the amphiploids derived from the cross between the wild barley Hordeum chilense and durum wheat. Tritordeums are characterised by higher yellow pigment content in their seeds than their durum wheat progenitors due to certain H. chilense genes located on the a arm of chromosome 7H(ch). In this work a candidate gene approach based on the phytoene synthase gene (PSY) was followed to investigate whether PSY1 may be responsible for the high carotenoid content in tritordeum and to develop a diagnostic marker for H. chilense PSY. This gene codes for the first step in the carotenoid biosynthetic pathway. It was first demonstrated that PSY is duplicated in H. chilense, Triticum urartu, and durum wheat ( PSY1 and PSY2), and subsequently a diagnostic cleaved amplified polymorphism ( CAP) marker able to differentiate between H. chilense and durum wheat PSY1 was developed. Using this CAP marker and a set of H. chilense-common wheat addition lines it was found that PSY1 is located on the alpha arm of chromosome 7H(ch), where the gene(s) for yellow pigment content are located. PSY1 is located on chromosomes 7A and 7B of durum wheat as demonstrated using Langdon substitution lines. Furthermore, synteny between rice and wheat indicates that PSY1 should be located on the long arms of chromosomes 7A and 7B, in agreement with QTL data for yellow pigment content. Together, these results suggest that PSY1 may be a good candidate gene for further work with yellow pigment content in both durum wheat and tritordeum. In addition, the diagnostic CAP marker developed will be used in our breeding program to transfer H. chilense genes to durum wheat, to evaluate their potential for durum wheat improvement.