Abstract
Ogura cytoplasmic male sterility (CMS) and its corresponding nuclear fertility restorer gene, Rfo, have been introduced from radish to Brassica species by interspecific crosses. Rfo restores male fertility by altering the translational expression of Orf138, a mitochondrial gene, whose expression results in the male sterile phenotype. This system has been extensively investigated and breeding restorer lines for the Ogura CMS has become a major objective for hybrid seed production in many canola breeding programs. In this study, we have sequenced genomic clones of Rfo amplified from a canola restorer line R2000, licensed from INRA, France, and a Dow AgroSciences non-restorer line Nexera 705 using primers designed from the radish Rfo sequence (GenBank accession AJ550021). Sequence alignment revealed three homologous sequences of Rfo. Two of the sequences were present in both R2000 and Nexera 705 but the third one was present only in R2000. These results suggested that the first two sequences could be the homoeologous sequences of Rfo already existing in the canola genome and the third one could be the radish Rfo introduced into canola. Based on the sequence differences between the restorer and non-restorer lines, Rfo allele-specific PCR markers were developed. We also developed a high throughput, Rfo allele-specific Invader® assay through Third Wave Technologies. Linkage analysis revealed a co-segregation between the allele-specific marker and the phenotypes for fertility restoration. This allele-specific marker has been mapped in the linkage group N19 and proved to be very useful for direct selection of Rfo alleles for fertility restoration during marker-assisted introgression of the Ogura restorer for hybrid development in canola.
Similar content being viewed by others
References
Bannerot H, Boulidard L, Cauderon Y, Tempé J (1974) Transfer of cytoplasmic male sterility from Raphanus sativus to Brassica oleracea. Proc RUCARPIA Crop Section Cruciferae 25:52–54
Bellaoui M, Grlon M, Pelletier G, Budar F (1999) The restorer Rfo gene acts post-translationally on the stability of the ORF138 Ogura CMS-associated protein in reproductive tissues of rapeseed cybrids. Plant Mol Biol 40:893–902. doi:10.1023/A:1006223908044
Bentolila S, Alfonso AA, Hanson MR (2002) A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants. Proc Natl Acad Sci USA 99:10887–10892. doi:10.1073/pnas.102301599
Bonhomme S, Budar F, Lancelin D, Small I, Defrance MC, Pelletier G (1992) Sequence and transcript analysis of the Nco2.5 Ogura specific fragment correlated with cytoplasmic male-sterility in Brassica cybrids. Mol Gen Genet 235:340–348. doi:10.1007/BF00279379
Brown GG, Formanova N, Jin H, Wargachuk R, Dendy C, Patil P et al (2003) The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J 35:262–272. doi:10.1046/j.1365-313X.2003.01799.x
Chase CD (2006) Genetically engineered cytoplasmic male sterility. Trends Pharmacol Sci 11:7–9. doi:10.1016/j.tplants.2005.11.003
de Arruda M, Lyamichev VI, Eis PS, Iszczyszyn W, Kwiatkowski RW, Law S et al (2002) Invader® technology for DNA and RNA analysis: principles and applications. Expert Rev Mol Diagn 2:487–496. doi:10.1586/14737159.2.5.487
Delourme R, Bouchereau A, Hubert N, Renard M, Landry BS (1994) Identification of RAPD markers linked to a fertility restorer gene for the Ogura radish cytoplasmic male sterility of rapeseed (Brassica napus L.). Theor Appl Genet 88:741–748. doi:10.1007/BF01253979
Delourme R, Foisset N, Horvais R, Barret P, Champagne G, Cheung WY et al (1998) Characterization of the radish introgression carrying the Rfo restorer gene for the Ogu-INRA cytoplasmic male sterility in rapeseed (Brassica napus L.). Theor Appl Genet 97:129–134. doi:10.1007/s001220050876
Desloire S, Gherbi H, Laloui W, Marhadour S, Clouet V, Catolico L et al (2003) Identification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Rep 4:588–594. doi:10.1038/sj.embor.embor848
Giancola S, Marhadour S, Desloire S, Clouet V, Falentin-Guyomarc’h H, Laloui W, Falentin C, Pelletier G, Renard M, Bendahmane A, Delourme R, Budar F (2003) Characterization of a radish introgression carrying the Ogura fertility restorer gene Rfo in rapeseed, using the Arabidopsis genome sequence and radish genetic mapping. Theor Appl Genet 107:1442–1451. doi:10.1007/s00122-003-1381-2
Hanson MR, Bentolila S (2004) Interactions of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16:S154–S169. doi:10.1105/tpc.015966
Hu X, Sullivan-Gilbert M, Gupta M, Thompson SA (2006) Mapping of the loci controlling oleic and linolenic acid contents and development of fad2 and fad3 allele-specific markers in canola (Brassica napus L.). Theor Appl Genet 113:497–507. doi:10.1007/s00122-006-0315-1
Koizuka N, Imai R, Fujimoto H, Hayakawa T, Kimura Y, Kohno-Murase J et al (2003) Genetic characterization of a pentatricopeptide repeat protein gene, orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant J 34:407–415. doi:10.1046/j.1365-313X.2003.01735.x
Kosambi DD (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175
Lyamichev V, Neri B (2003) Invader® assay for SNP genotyping. Methods Mol Biol 212:229–240
Ogura H (1968) Studies on the new male sterility in Japanese radish, with special reference to the utilization of this sterility towards the practical raising of hybrid seeds. Mem Fac Agric Kagoshima Univ 6:38–78
Pelletier G, Primard C, Vedel F, Chétrit P, Rémy R, Rousselle P et al (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191:244–250. doi:10.1007/BF00334821
Primard-Brisset C, Poupard JP, Horvais R, Eber F, Pelletier G, Remard M et al (2005) A new recombined double low restorer line for the Ogu-INRA cms in rapeseed (Brassica napus L.). Theor Appl Genet 111:736–746. doi:10.1007/s00122-005-2059-8
Van Ooijen JW, Voorrips RE (2001) JoinMap® Version 3.0: Software for the calculation of genetic linkage maps. Plant Research International, Wageningen
Acknowledgements
Authors would like to thank Dr. Van Ripley and Yonghe Bai for their critical review of the manuscript and valuable comments.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hu, X., Sullivan-Gilbert, M., Kubik, T. et al. Mapping of the Ogura fertility restorer gene Rfo and development of Rfo allele-specific markers in canola (Brassica napus L.). Mol Breeding 22, 663–674 (2008). https://doi.org/10.1007/s11032-008-9207-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11032-008-9207-1