Development of markers linked to Diuraphisnoxia resistance in wheat using a novel PCR-RFLP approach

Through random amplified polymorphic DNA (RAPD) analysis we identified a putative marker linked to the Dn5 resistance gene. This marker was converted to a more reliable sequence-characterised-amplified regions (SCAR) marker. The initial SCAR marker amplified the correct amplification product but failed to discern between the susceptible and resistant individuals. Hence, it was utilised to sequence the internal fragment. All nested primers designed from the internal sequences were also unable to produce any polymorphism between the susceptible and resistant cultivars. Restriction digests were then performed on these fragments, and the restriction enzyme EcoRI was able to discern between the susceptible and resistant F₂ individuals of the Dn5 population. This granted one marker amplified with the internal SCAR primer set OPF14₁₀₈₃ the ability to differentiate between parental individuals carrying the Dn5 genes. This marker was tested in a segregating F₂ population carrying the Dn5 resistance gene and proved able to differentiate between the segregating individuals. This marker may prove useful in marker assisted selection (MAS), although performing restriction digests may hamper the throughput of a high number of samples. Received: 4 August 1999 / Accepted: 27 August 1999