Location of markers of aluminium tolerance genes on rye chromosomes (Secale cereale L.)

Iwona Wiśniewska

Plant Breeding and Acclimatisation Institute, Radzików, 05-870 Błonie, Poland (Poland)

Andrzej Rafalski

Plant Breeding and Acclimatisation Institute, Radzików, 05-870 Błonie, Poland (Poland)

Abstract

The aim of presented work was to identify of PCR amplifiedDNAfragments differentiating aluminium tolerant and sensitive forms of rye and to locate the markers on rye chromosomes. For identification of markers, the PCR system with semi-specific primers targeting intron-exon sequences of plant genes was applied. The modified method of bulked segregant analysis was used. The pooled DNAs of two or three F₂ segregating populations were screened together with DNA of their parental inbred lines. Potential marker of tolerance gene was located on rye chromosomes using wheat/rye (Chinese Spring/Blanco) additional lines. The specific probes obtained from DNA fragments differentiating sensitive and tolerant forms of rye were hybridized to PCR amplified DNA fragments of sensitive and tolerant forms of rye and the set of wheat/rye addition lines. Independently of the method of digoxygenin labelling (primer extension or Taq polymerase reaction), the probes obtained showed similar hybridization patterns. The results of hybridisation of 21 probes prepared from 12 DNA fragments confirmed connection of selected DNA fragments with to aluminium tolerance or sensitivity. Most of these DNA fragments originated from tolerant forms of rye. Using this method it was possible to locate eight DNA fragments on rye chromosomes. Three DNA fragments hybridised to chromosome 4R, two DNA fragments to chromosome 6R and single DNA fragments to chromosomes 1R, 2R and 3R. Four DNA fragments indicating clear relationship with character studied were not located on particular chromosomes using this set of wheat/rye addition lines. Hybridisation of probes prepared from four DNA fragments revealed length polymorphism. Probes prepared from two DNA fragments were characterised as dominant markers. In other cases the type of marker (dominant/codominat) was not fully documented.

Keywords:

aluminium tolerance, chromosomal location, DNA-DNA hybridisation, PCR markers, wheat/rye addition lines