Molecular tagging of the male fertility restorer gene for the 501-8S cytoplasmic male sterility in rapeseed (Brassica napus L.)
Haohua He A , Liang Xu B , Xiaosong Peng A , Guangsheng Yang C , Changlan Zhu A , Zunwen Liu D and Guoyou Ye E FA Crop Breeding Research Institute, Jiangxi Agricultural University, Nanchang 330045, China.
B Qinghai Academy of Agriculture and Forestry, Xining 810016, China.
C National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China.
D Yichun Institute of Agricultural Sciences, Yichun 336000, China.
E Department of Primary Industries Victoria, and Molecular Plant Breeding Cooperative Research Centre, 1 Park Drive, Bundoora, Vic. 3086, Australia.
F Corresponding author. Email: guoyou.ye@dpi.vic.gov.au
Australian Journal of Agricultural Research 58(8) 753-758 https://doi.org/10.1071/AR06369
Submitted: 22 November 2006 Accepted: 23 April 2007 Published: 30 August 2007
Abstract
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 F1 and F2, BC1, and F3 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 F2 population of the cross 501-8S × Yuyou1 was used as a mapping population to map the Rf gene. A combination of bulked segregant analysis (BSA) and amplified 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, EA02MG03-260, is only 0.4 cM from the Rf gene. The EA02MG03-260 marker was converted to a dominant sequence characterised amplified region (SCAR) marker (SCARE2M3-214). Amplification using this locus-specific primer generated specific bands with male fertile plants when tested using the mapping population. Specific amplification of SCARE2M3-214 was also detected in all 3 male sterile plants and their F1 hybrids, with 5 restorer lines used for verification. Thus, SCARE2M3-214 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.
Additional keywords: AFLP, fertility restoration, hybrids, oilseed rape.
Acknowledgments
We are grateful for financial support from the National Natural Science Foundation of China and the Department of Science and Technology of Jiangxi Province, China. We also thank Dr Maarten van Ginkel (Department of Primary Industries, Vic., Australia) for critical editing of the manuscript.
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