Mapping of the Ogura fertility restorer gene Rfo and development of Rfo allele-specific markers in canola (Brassica napus L.)

Hu, Xiaokang; Sullivan-Gilbert, Mandy; Kubik, Tom; Danielson, Jason; Hnatiuk, Nathan E.; Marchione, Wesley; Greene, Thomas W.; Thompson, Steven A.

doi:10.1007/s11032-008-9207-1

Cited by 29 publications

(18 citation statements)

References 16 publications

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“…Based on the sequence differences between the restorer and non-restorer lines in canola, Hu et al (2008) confirmed that Rfo is located on linkage group N19 the Invader assay and simplesequence repeat analysis. Yasumoto et al (2008a) performed PCR-restriction fragment length polymorphism analysis to investigate the distribution of the Rf gene (orf687) for Ogura male-sterile cytoplasm in Japanese wild radish.…”

Section: Discussionmentioning

confidence: 66%

Molecular characterization of the Rs-Rf 1 gene and molecular marker-assisted development of elite radish (Raphanus sativus L.) CMS lines with a functional marker for fertility restoration

et al. 2012

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Cytoplasmic male sterility (CMS) is a maternally inherited trait that is widely utilized in hybrid seed production. In this study, the Rs-Rf 1 gene and its allele, the Rs-rf 1 gene, were isolated from a radish CMS restorer line and maintainer, respectively. The Rs-Rf 1 gene was identified as a member of the pentatricopeptide repeat family. The results of reverse transcription (RT)-PCR and quantitative RT-PCR analyses of the floral buds at different development stages were used to characterize the expression features of Rs-Rf 1 and orf138, a unique transcribed gene in the mitochondrion for its role in determining male sterility in the cytoplasm. orf138 was upregulated in the CMS line and F 1 hybrid at three bud developmental stages, including meiosis and the tetroid and microspore stages, but it was not expressed in the maintainer and restorer line, suggesting that the Rs-Rf 1 gene was not able to block orf138 transcription. As in the F 1 plant, the transcription of Rs-Rf 1 was gradually up-regulated in the restorer line with progressive stages of bud development, but it was not transcribed in the CMS line and maintainer. Different restriction sites for SspI between the allele of Rs-Rf 1 and Rs-rf 1 were also identified, and a guideline for using the functional marker to distinguish the genotypes of individuals with Rs-Rf 1 was developed. Due to its co-dominant character, this marker could be used to discriminate the heterozygous and homozygous fertility-restoring allele. Using this functional marker, we established a system of molecular marker-assisted development of elite CMS lines in radish. This system will contribute greatly to candidate maintainer identification and accelerate the process of elite CMS line development in late-bolting radish breeding programs.

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Section: Discussionmentioning

confidence: 66%

Molecular characterization of the Rs-Rf 1 gene and molecular marker-assisted development of elite radish (Raphanus sativus L.) CMS lines with a functional marker for fertility restoration

et al. 2012

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“…To achieve complete stability of the Rfk1 gene, also exploited in practical breeding, the integration of the gene to the A genome chromosomes of turnip rape may be required. The Rfo gene, which is stably integrated in the oilseed rape C genome (Hu et al 2008;Feng et al 2009), is now used for hybrid seed production (Budar et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Size and location of radish chromosome regions carrying the fertility restorer Rfk1 gene in spring turnip rape

et al. 2012

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In spring turnip rape (Brassica rapa L. spp. oleifera), the most promising F1 hybrid system would be the Ogu-INRA CMS/Rf system. A Kosena fertility restorer gene Rfk1, homolog of the Ogura restorer gene Rfo, was successfully transferred from oilseed rape into turnip rape and that restored the fertility in female lines carrying Ogura cms. The trait was, however, unstable in subsequent generations. The physical localization of the radish chromosomal region carrying the Rfk1 gene was investigated using genomic in situ hybridization (GISH) and bacterial artificial chromosome-fluorescence in situ hybridization (BAC-FISH) methods. The metaphase chromosomes were hybridized using radish DNA as the genomic probe and BAC64 probe, which is linked with Rfo gene. Both probes showed a signal in the chromosome spreads of the restorer line 4021-2 Rfk of turnip rape but not in the negative control line 4021B. The GISH analyses clearly showed that the turnip rape restorer plants were either monosomic (2n=2x=20+1R) or disomic (2n=2x=20+2R) addition lines with one or two copies of a single alien chromosome region originating from radish. In the BAC-FISH analysis, double dot signals were detected in subterminal parts of the radish chromosome arms showing that the fertility restorer gene Rfk1 was located in this additional radish chromosome. Detected disomic addition lines were found to be unstable for turnip rape hybrid production. Using the BAC-FISH analysis, weak signals were sometimes visible in two chromosomes of turnip rape and a homologous region of Rfk1 in chromosome 9 of the B. rapa A genome was verified with BLAST analysis. In the future, this homologous area in A genome could be substituted with radish chromosome area carrying the Rfk1 gene.

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“…Although suitable molecular markers for the Rfo gene in Brassica napus have been developed [5,7,8], their utilisation in DH Rf line production requires cultivation of DH regenerants up to a stage when there is a sufficiently large leaf area for DNA isolation and subsequent analyses. A method that would enable the non-destructive identification of genotypes carrying the Rfo gene at early stages of microspore derived embryos would be of great benefit in terms of significantly reducing the amount of cultivated DH regenerants, and thus decreasing workload, culture space, time and cost.…”

Section: Introductionmentioning

confidence: 99%

Non-destructive in vitro selection of microspore-derived embryos with the fertility restorer gene for CMS Ogu-INRA in winter oilseed rape (Brassica napus L.)

Havlíčková

Klíma

Pribylova

et al. 2015

Electronic Journal of Biotechnology

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We have bred low glucosinolate (GSL) winter oilseed rape lines carrying the fertility restorer for the CMS Ogu-INRA system. The original restorer line BO20 contained 31μmol/g GSL in seeds, but by crossing this line with various low GSL CMS lines, followed by repeated selection of fertile segregants, we were able to obtain fertile lines with a mean GSL content in seeds of 11.8 μmol/g. This result confirmed that the gene(s) controlling the GSL content are not closely linked to the fertility restorer gene. The results confirm, that the SCAR marker SG34 is closely associated with the fertility restoring allele, and facilitates so the selection of fertile segregants; however, the marker is unable to distinguish between the homozygous RfRf and the heterozygous Rfrf genotypes.

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Mapping of the Ogura fertility restorer gene Rfo and development of Rfo allele-specific markers in canola (Brassica napus L.)

Cited by 29 publications

References 16 publications

Molecular characterization of the Rs-Rf 1 gene and molecular marker-assisted development of elite radish (Raphanus sativus L.) CMS lines with a functional marker for fertility restoration

Molecular characterization of the Rs-Rf 1 gene and molecular marker-assisted development of elite radish (Raphanus sativus L.) CMS lines with a functional marker for fertility restoration

Size and location of radish chromosome regions carrying the fertility restorer Rfk1 gene in spring turnip rape

Non-destructive in vitro selection of microspore-derived embryos with the fertility restorer gene for CMS Ogu-INRA in winter oilseed rape (Brassica napus L.)

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