Development of a DNA marker for Fusarium wilt resistance in chickpea

Fusarium wilt caused by Fusarium oxysporum Schlechtend.:Fr. f. sp. ciceris (Padwick) Matuo & K. Sato is the most widely spread soilborne disease of chickpea (Cicer arietinum L.). To advance our understanding of the genetics of wilt resistance and aid chickpea breeding programs, we developed a set of F6 recombinant inbred lines (RILs) between Fusarium wilt susceptible (C-104) and resistant (WR-315) parents. Prior screening of selected F3 plants identified two primers (UBC-170 and CS-27) that produced random amplified polymorphic DNA (RAPD) markers associated with Fusarium wilt race 1 resistance. Analysis of the RILs with these primers yielded an estimate of 7% recombination between the two markers and the locus for wilt resistance, and 6% recombination between the loci corresponding to the two RAPD markers. The DNA fragments were cloned and sequenced in order to construct primers that would amplify only the markers of interest. Primer pair CS-27F/CS-27R amplified a fragment linked to the allele for susceptibility to race 1 of Fusarium wilt and thus constitute allele specific associated primers (ASAPs), whereas UBC-170F/UBC-170R produced a single band for both resistant and susceptible genotypes, thus demonstrating locus specificity rather than allele specificity. The use of markers generated by the RAPD or ASAP approaches can facilitate the introgression of resistance genes into susceptible lines and expedite the screening of chickpea germplasm resources and will be useful in extending the genetic map of chickpea.