Identification of AFLP markers linked to fertility restorer genes for tournefortii cytoplasmic male-sterility system in Brassica napus

The tournefortii cytoplasmic male-sterility system is being used as a method of pollination control to develop hybrids in Brassica napus. Genetic analyses have indicated that two dominant genes, one major (Rft1) and another minor (Rft2), were required to achieve complete fertility restoration. Though the major gene (Rft1) can cause complete fertility restoration on its own, its expression was significantly enhanced in the presence of the minor gene (Rft2). In the absence of Rft1, Rft2 caused only partial fertility restoration. We used a pair of near-isogenic lines (NILs), differing for the presence/absence of Rf genes, to identify AFLP markers linked to fertility restorer genes. A total of 64 EcoRI/MseI primer combinations were surveyed which produced 3,225 bands, of which 19 (0.006%) were polymorphic between parental NILs. Primer combinations which led to the identification of polymorphic bands present in fertile parental NILs were used for assaying a mapping population of 70 F₂ plants for determining the segregation pattern of markers. Initial screening resulted in the identification of five AFLP markers. The recombination analyses of these AFLP markers revealed that at least two (EACC/MCTT₁₀₅, EAAG/MCTC₈₀) were present in the same linkage group along with the Rf loci. Marker EACC/MCTT₁₀₅ was separated from the major gene (Rft1) by a distance of 18.1 cM, while it was 33.2 cM away from the minor fertility restorer gene (Rft2). Another marker EAAG/MCTC₈₀ was also located adjacent to Rft1 at a distance of 18.1 cM, but on other side. Identification of flanking markers (EACC/MCTT₁₀₅, EAAG/MCTC₈₀) for the major fertility restorer gene (Rft1) provides a crucial component for marker-assisted selection and map-based cloning of the restorer genes, and can hence be used to construct elite restorer genotypes.