Using a gel‐free PCR‐ELISA for the molecular identification of wheat genotypes carrying wheat–rye translocations

Abstract

Current techniques to identify wheat lines possessing the 1RS chromosome are generally unsuitable in relation to the speed and cost needs in modern wheat breeding programmes. A gel-free, direct amplicon capture PCR-ELISA assay was developed and evaluated, aiming at speeding the identification of wheat genotypes possessing the 1RS chromosome arm in breeding programmes. The chosen target sequence was the repetitive, interspersed rye-specific element RIS-1. Primers were end-labelled with digoxigenin and biotin, and amplicons captured on to straptavidine-coated microplates. Subsequent immunodetection of the digoxigenin moiety readily distinguished 1RS from non-1RS control genotypes tested. When a nursery consisting of 120 winter and spring wheat lines was screened by PCR-gel electrophoresis and the PCR-ELISA, a perfect agreement between both techniques was observed. Test robustness, as measured by the tolerance to variations in DNA input, was better for PCR-ELISA than PCR-gel electrophoresis. In conclusion, a simple, robust, fast and scalable technique for the detection of 1RS chromosome carriers in wheat breeding programmes is now available.

Testing of rye-specific markers located on 1RS chromosome and distribution of 1AL.RS and 1BL.RS translocations in Turkish wheat (Triticum aestivum L., T. durum Desf.) varieties and landraces

“…Globally, numerous wheat cultivars, particularly carrying the 1BL.1RS wheat-rye translocation, have been released (Rabinovich 1998). Therefore, it is of paramount importance for every wheat breeding program to know the inheritance and distribution of these translocations, Several methods such as C-banding, in situ hybridization, ELISA, protein electrophoresis, disease response tests and DNA markers have been used to detect rye chromosome arms or fragment in the wheat genome (Gill and Kimber 1977;Zeller and Fuchs 1983;Heslop-Harrison et al 1990;Weng et al 2007;Zuniga et al 2008). DNA analysis, based on PCR, is one of the main approaches to identify 1AL.1RS and 1BL.1RS translocations in wheat background.…”