Molecular probes for detection of wheat chromosomal fragments in rye.

Andrzej Rafalski

Department of Plant Biochemistry and Physiology, Plant Breeding and Acclimatization Institute, Radzików, 05-870 Błonie, Poland (Poland)

Iwona Wiśniewska

Department of Plant Biochemistry and Physiology, Plant Breeding and Acclimatization Institute, Radzików, 05-870 Błonie, Poland (Poland)

Teresa Sikora

National Centre for Plant Genetic Resources, Plant Breeding and Acclimatization Institute, Radzików, 05-870 Błonie, Poland (Poland)

Bogusław Łapiński

National Centre for Plant Genetic Resources, Plant Breeding and Acclimatization Institute, Radzików, 05-870 Błonie, Poland (Poland)

Abstract

Twelve DNA fragments amplified on templates of Triticum urartu, Aegilops speltoides and Aegilops squarrosa were separated, reamplified and used as digoxigenin-labelled probes. The species and genome specificity of probes was evaluated by Southern dot-blot hybridisation to Triticum durum, the donors of wheat genomes, rye and rye with introgressed wheat chromosome fragments. In comparison to labelled genomic DNA of T. durum, the probes obtained by labelling PCR-amplified fragments exhibited higher specificity to wheat genomes. Under the applied hybridisation conditions all probes showed different degree of cross-hybridisation to rye DNA. Some probes indicated quantitative difference in their specificity to wheat genomes, but generally the intensity of hybridisation to the genomes A, S and D was independent of the origin of an amplified fragment. Selected probes, used in dot-blot hybridisation system together with genomic DNA of T. durum, may increase the sensitivity of screening wheat chromosomal fragments introgressed to rye.

Keywords:

introgression, PCR, rye, Southern blotting, Triticum durum probe, wheat, wheat specific probes