Molecular tagging of the male fertility restorer gene for the 501-8S cytoplasmic male sterility in rapeseed (Brassica napus L.)

He, H.X.,,, G.Z.u, C.L.u, Z.Y., G.

Australian journal of agricultural research 8(8): 753-758


ISSN/ISBN: 0004-9409
DOI: 10.1071/ar06369
Accession: 016410288

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The cytoplasmic male sterile (CMS) system has been successfully used to explore heterosis in rapeseed ( Brassica napus L.). A newly developed male sterile line (501-8S) was characterised for its male fertility response to temperature and photoperiod, and the inheritance of fertility restoration. Segregation analysis using F-1 and F-2, BC1, and F-3 populations of the crosses between the 501-8S and fertile lines of B. napus revealed that fertility restoration was conferred by a dominant nuclear gene (Rf). The F-2 population of the cross 501-8S x Yuyou1 was used as a mapping population to map the Rf gene. A combination of bulked segregant analysis ( BSA) and amplifed fragment length polymorphism (AFLP) methodology was used to identify putative markers linked to the Rf gene. Twenty-nine of the 1280 primer combinations tested revealed polymorphism between the 2 extreme bulks. Further testing of these primer combinations in individual plants identified 5 AFLP markers tightly linked to the Rf gene with a map distance of less than 5.0 cM. All 5 markers were on one side of the restoration gene in the coupling phase. The closest marker, EA(02)MG(03)-260, is only 0.4 cM from the Rf gene. The EA(02)MG(03)-260 marker was converted to a dominant sequence characterised amplified region ( SCAR) marker (SCAR(E2M3)-214). Amplification using this locus-specific primer generated specific bands with male fertile plants when tested using the mapping population. Specific ampli. cation of SCAR(E2M3)-214 was also detected in all 3 male sterile plants and their F-1 hybrids, with 5 restorer lines used for veri. cation. Thus, SCAR(E2M3)-214 will be very useful for the development of new restorer lines by the transfer of the Rf gene into other breeding lines. It can also be used for isolating the Rf gene by means of map-based cloning.